Abstract:ObjectiveTo explore the mechanism of Fuzi Lizhongwan alleviating the damage of chemotherapy-induced peripheral neuropathy (CIPN) mice caused by cisplatin based on mitogen-activated protein kinase (MAPK) signaling pathway.MethodA total of 40 female KM mice were randomized into blank group (distilled water, ig), model group (distilled water, ig), Fuzi Lizhongwan group (3.5 g·kg-1, ig), and aspirin group (0.026 g·kg-1, ig). Cisplatin (3 mg·kg-1, ip, 5 days) was used to induce CIPN in mice. Administration began while modeling and lasted 12 days. The general conditions and behaviors of mice were observed. After the last administration, samples were collected. Pathological changes of the soles were observed based on hematoxylin-eosin (HE) staining. Biochemical assay was employed to determine the levels of serum superoxide dismutase (SOD), hydrogen peroxide (H2O2), malondialdehyde (MDA), and nitric oxide (NO), enzyme-linked immunosorbent assay (ELISA) the content of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and glutathione peroxidase-3 (GPX-3) in kidney tissue, and Western blotting the expression of extracellular signal-regulated kinase1/2 (ERK1/2), phosphorylated-ERK1/2 (p-ERK1/2), p38 MAPK, and phosphorylated-p38 MAPK (p-p38 MAPK) in kidney tissue.ResultCompared with the blank group, model group demonstrated obvious pathological damage on the soles, hyperkeratosis of the epidermis with a basketweave pattern, atrophy of stratum spinosum, reduction of cells, and intracellular edema. Compared with the model group, Fuzi Lizhongwan significantly alleviated the pathological damage of the skin tissue of the soles. The model group showed lower body weight, mechanical pain threshold, thermal pain threshold (P<0.01), and SOD activity (P<0.05), higher content of H2O2, MDA, and NO (P<0.01), and higher expression of IL-6, IL-1β, and TNF-α (P<0.01) than the blank group. Fuzi Lizhongwan group demonstrated higher body weight, mechanical pain threshold, thermal pain threshold (P<0.01), and SOD activity (P<0.05), lower content of H2O2, MDA, and NO (P<0.05), and lower expression of IL-6, IL-1β, and TNF-α (P<0.01) than the model group. The expression of ERK1/2, p-ERK1/2, p38 MAPK, and p-p38 MAPK increased significantly (P<0.01) in the model group compared with that in the blank group, while the expression decreased significantly (P<0.01) in the Fuzi Lizhongwan group compared with that in the model group.ConclusionFuzi Lizhongwan can relieve the neurological injury of cisplatin-induced CIPN mice and increase the pain threshold of mice, possibly by regulating the MAPK signaling pathway and inhibiting inflammatory response and oxidative stress.
Abstract:ObjectiveTo explore the mechanism of Liu Junzitang in preventing and treating muscle atrophy in mice with lung cancer cachexia based on the signal transducer and activator of transcription 3(STAT3)/ubiquitin proteasome pathway in vivo.MethodForty C57BL/6 mice aged six weeks were randomly divided into a blank group, a model group, a Liu Junzitang group, an inhibitor group (stattic group),and a Liu Junzitang + inhibitor group (combination group), with eight mice in each group. The cachectic muscle atrophy model was induced by subcutaneous inoculation of Lewis lung cancer cell line under the right anterior armpit in mice except those in the blank group. On the 8th day after subcutaneous inoculation, the mice in the corresponding groups received Liu Junzitang (9.56 g·kg-1·d-1) by gavage and intraperitoneal injection of stattic [25 mg·kg-1·(2 d)-1]. After three weeks of drug intervention, the body weight and gastrocnemius muscle weight were recorded. Hematoxylin-eosin (HE) staining was used to observe the pathological changes and cross-sectional area of gastrocnemius muscle fibers in mice. Western blot was used to detect the expression of phosphorylated-STAT3 (p-STAT3), STAT3, muscle atrophy F-box (MAFbx), and muscle RING finger protein 1 (MuRF1) in the gastrocnemius muscle. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of STAT3, MAFbx, and MuRF1 in the gastrocnemius muscle.ResultCompared with the blank group, the model group showed lightened body and the gastrocnemius muscle, reduced cross-sectional area of gastrocnemius muscle fibers, and increased protein expression of p-STAT3, STAT3, MAFbx, and MuRF1 and mRNA expression of STAT3, MuRF1, and MAFbx in the gastrocnemius muscle (P<0.05). Compared with the model group, the Liu Junzitang group showed increased body weight, gastrocnemius muscle weight, and cross-sectional area of gastrocnemius muscle fibers (P<0.05), and decreased protein expression of p-STAT3, STAT3, MuRF1, MAFbx, and mRNA expression of STAT3 and MAFbx in gastrocnemius muscle (P<0.05). Compared with the model group, the inhibitor group showed increased body weight and cross-sectional area of gastrocnemius muscle fibers (P<0.05), and reduced protein expression of p-STAT3, STAT3, MuRF1, MAFbx, and mRNA expression of STAT3 and MAFbx in gastrocnemius muscle (P<0.05). Compared with the model group, the combination group showed increased body weight and gastrocnemius muscle weight (P<0.05),and decreased protein expression of p-STAT3, STAT3, MuRF1, and mRNA expression of STAT3 and MAFbx in the gastrocnemius muscle (P<0.05). Compared with the Liu Junzitang group, the stattic group and the combination group showed reduced expression of p-STAT3 protein in the gastrocnemius muscle (P<0.05).ConclusionLiu Junzitang can prevent and treat muscle atrophy in mice with lung cancer cachexia, and its mechanism may be associated with the protein and mRNA expression related to the STAT3-mediated ubiquitin proteasome pathway.
Keywords:lung cancer cachexia;muscle atrophy;Liu Junzitang;signal transducer and activator of transcription 3(STAT3);ubiquitin proteasome
Abstract:ObjectiveTo investigate the effect of Shaoyaotang on diarrhea, inflammation, and intestinal flora in rats with dampness-heat diarrhea and explore the mechanism of therapeutic principle "treating incontinent syndrome with dredging method" of Shaoyaotang.MethodThe dampness-heat diarrhea model was induced by high temperature, high humidity, high sugar and fat diet, and pathogenic factors. The rats were divided into normal group, model group (normal saline), Shaoyaotang group (5.62 g·kg-1), Rhei Radix et Rhizoma (RRER)-free Shaoyaotang group (5.15 g·kg-1), and RRER group (0.01 g·kg-1). The rats were treated correspondingly for five days, twice a day in the morning and evening. The diarrhea index was used to evaluate the antidiarrheal effect of each group three hours after the administration in the evening. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-2, and IL-6 in the serum were detected by enzyme-linked immunosorbent assay (ELISA) three hours after the last administration. The structure of intestinal flora in feces was characterized by 16sDNA.ResultCompared with the model group, the Shaoyaotang group, the RRER-free Shaoyaotang group, and the RRER group showed reduced diarrhea index (P<0.01), with the onset rates ranking as the Shaoyaotang group>the RRER-free Shaoyaotang group>the RRER group. Those three groups with drug intervention all showed decreased levels of inflammatory factors (P<0.01), especially the Shaoyaotang group, and no significant difference was observed between the RRER group and the RRER-free Shaoyaotang group. The abundance of pathogenic bacteria and conditioned pathogens (e.g. Escherichia-Shigella, Prevotella, Enterorhabdus, and Bacteroides) was reduced and the proliferation of probiotics (such as Ruminococcus, Turicibacter, and Lachnospiraceae) was increased in the groups with drug intervention (P<0.01). For the structure of intestinal flora, the RRER group and the Shaoyaotang group were close to the normal group, and the RRER-free Shaoyaotang group was different from the other three groups (P<0.01).ConclusionShaoyaotang can improve the outcome of rats with dampness-heat diarrhea through anti-inflammation and regulation of intestinal flora disorders. RRER in the prescription plays a key role in reducing the abundance of harmful bacteria and promoting the proliferation of probiotics, which is the key of Shaoyaotang in promoting the re-balance of intestinal flora. It also confirms the scientificity of treating dampness-heat diarrhea with RRER following the therapeutic principle "treating incontinent syndrome with dredging method".
Keywords:Shaoyaotang;dampness-heat diarrhea;intestinal flora;16sDNA;Rhei Radix et Rhizoma
Abstract:ObjectiveTo observe the effects of Qingzao Jiufeitang on the expression of adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), and UNC-51-like kinase 1 (ULK1) in lung cancer cells after the application of AMPK inhibitor (compound C).MethodMale C57BL/6J mice were randomly divided into a model group, a cyclophosphamide (CTX) group (50 mg·kg-1), a Qingzao Jiufeitang group (11 g·kg-1), an AMPK inhibitor group (10 mg·kg-1), and a Qingzao Jiufeitang combined with AMPK inhibitor group (combination group) (11 g·kg-1+10 mg·kg-1). Lewis lung cancer cells were subcutaneously injected into the right axilla to induce a tumor-bearing model. 24 hours after modeling, the mice in the CTX group were intraperitoneally injected once every other day for seven times in total. The mice in the AMPK inhibitor group and the combination group received intraperitoneal injection of compound C, once a day for 14 days. The mice in the Qingzao Jiufeitang group and the combination group were administered orally at the set dose for 14 days before and after modeling. At the end of the experiment, the mice in each group were sacrificed. The tumor-bearing tissues were collected, and the tumor weight of each group was counted. Transmission electron microscopy (TEM) was used to observe the formation of autolysosomes in lung cancer tissues of each group. Western blot was used to detect the protein expression of AMPK, phosphorylated AMPK (p-AMPK), mTOR, phosphorylated mTOR (p-mTOR), ULK1, phosphorylated ULK1 (p-ULK1), microtubule-associated protein 1 light chain 3B (LC3B), and p62. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung cancer in each group.ResultCompared with the model group, the Qingzao Jiufeitang group showed decreased tumor weight (P<0.01), the formation of autolysosomes under the electron microscope, increased protein expression of p-AMPK, p-ULK1, LC3B, LC3B-Ⅱ, and p-AMPK/AMPK, p-ULK1/ULK1, and LC3B-Ⅱ/LC3B-Ⅰratios (P<0.01, P<0.05), and reduced protein expression of p-mTOR, p62, and p-mTOR/mTOR ratio (P<0.05). Compared with the Qingzao Jiufeitang group, the combination group showed no autolysosomes formation under the electron microscope, decreased protein expression of p-AMPK, p-ULK1, LC3B, LC3B-Ⅱ, and p-AMPK/AMPK, p-ULK1/ULK1, LC3B-Ⅱ/LC3B-Ⅰ ratios (P<0.05, P<0.01), and increased p62 protein expression (P<0.05). HE staining results showed that the pathological changes of lung cancer tissues in the groups with drug intervention were improved compared with those in the model group.ConclusionQingzao Jiufeitang can promote the elevation of LC3B-Ⅱ and decrease the expression of p62 protein, thus inducing autophagy. The mechanism of autophagy initiation may be achieved by the AMPK/ULK1 pathway instead of the mediation by the AMPK/mTOR/ULK1 pathway.
Keywords:adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK);Qingzao Jiufeitang;lung cancer;proteins related to autophagy initiation
Abstract:ObjectiveTo observe the intervention of modified Sanrentang on the lipopolysaccharide-induced proliferation of rat glomerular mesangial cells, the phosphatidylinositol-3 kinase(PI3K)/protein kinase B(PKB/Akt)/nuclear factor kappa B(NF-κB) signaling pathway, and to investigate its mechanism in improving kidney inflammation in rats with immunoglobulin A nephropathy(IgAN).MethodThe 18 rats were divided into 3 groups by serum pharmacology method: normal group, high-dose and low-dose (20.70,10.35 g·kg-1·d-1) groups with 6 rats in each group. Modified Sanrentang high- and low-dose groups were intragastric with the corresponding solution of modified Sanrentang, and normal group was intragastric with equal volume of distilled water. After 5 days of intragastric administration, blood samples were collected to prepare drug-containing serum. Rat mesangial (HBZY-1) were divided into five groups of normal group, LPS 10 mg·L-1 in the model group, benazepril(50 μmol·L-1), modified Sanrentang high- and low-dose group. Preclude the use of methyl thiazolyl tetrazolium(MTT) method detect the proliferation activity of HBZY-1 cells, enzyme-linked immunosorbent assay(ELISA) was used to determine the content of each group type Ⅳ collagen(ColⅣ),Western blot and Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) were used to detect protein and mRNA expression levels of PI3K/Akt/NF-κB signaling pathway.ResultAs compared with the normal group, MTT assay showed that exposure to LPS significantly enhanced the proliferative activity, the ColⅣ was increased significantly of HBZY-1 cells(P<0.01), p-Akt, p-p65 was increased significantly (P<0.01). Compared with the model group, the proliferation and ColⅣ of rat chronic glomerulonephritis cells induced by LPS by inhibiting PI3K/Akt/NF-κB signaling pathway(P<0.01), and the phosphorylation of Akt was significantly inhibited(P<0.01), the expression levels of NF-κB p65 was reduced in modified Sanrentang high-dose group(P<0.01).ConclusionModified Sanrentang could inhibit cell proliferation and the content of ColⅣ in rat mesangial cells induced by LPS, and its mechanism might be related to suppression of PI3K/Akt signaling pathway.
Abstract:ObjectiveTo observe that effect of Ersi decoction on rats with rheumatoid arthritis (RA) induced by using the complete Freund's adjuvant emulsion containing bovine type Ⅱ collagenand and elucidate underlying menchanisms involving to inhibit inflammation and joint synovial angiogenesis.MethodThe rat model of RA was established by immune induction with complete Freund's adjuvant emulsion containing bovine type Ⅱ collagen. All male SD rats were randomly divided into blank group, RA model group, methotrexate group(1.0 mg·kg-1), and low-, medium- and high-dose group(30,15,7.0 g·kg-1·d-1)of Ersi decoction, with 8 rats in each group. Except the blank group, rats in the methotrexate group and Ersi decoction groups were given corresponding doses of methotrexate and Ersi decoction after establishment of RA induced by strengthen immunity,respectively,and those in the model group and blank group received normal saline of equivalent volume,once a day for 28 days. After the administration, the degree of joint swelling of rats in each group was analyzed by joint swelling volume and index. The small animal ultrasound imaging system was used to detect the score and area of synovial hyperplasia of knee joint in right lower limb of rats and hematoxylin-eosin(HE)staining to observe the histomorphological changes in joint synovium of rats. The levels of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were measured by enzyme-linked immunosorbent assay(ELISA). Immunohistochemistry was employed to analyze the expression of CD31 and vascular endothelial growth factor receptor 2(VEGFR2) in in joint synovium.ResultCompared with the blank group, the model group demonstrated significant increase in joint swelling volume and index, inflammatory cytokines including TNF-α and IL-1β in serum, the score and area of synovial hyperplasia of knee joint in right lower limb, obvious pathological changes in the synovium and the expression of CD31 and VEGFR2 in joint synovium. Medium and high-dose Ersi decoction significantly alleviated the pathological changes of synovium tissue, attenuated joint swelling volume and index and decreased the expression of CD31 and VEGFR2 in joint synovium as compared with the model group. Moreover, high-dose Ersi decoction showed significantly lower levels of TNF-α and IL-1β in serum, and the score and area of synovial hyperplasia of knee joint in right lower limb. But medium-dose Ersi decoction only showed lower levels of TNF-α and area of synovial hyperplasia of knee joint.ConclusionErsi decoction could reduce synovial inflammation and hyperplasia through inhibiting synovial angiogenesis in rats with RA induced by bovine type Ⅱ collagen for achieving the effect of reducing RA joint damage, which provides an important reference for anti-RA of Ersi decoction in clinical application.
Abstract:ObjectiveTo explore the effect and mechanism of Zhiwang decoction combined with methotrexate (MTX) against bone destruction in rats with collagen-induced arthritis (CIA).MethodMale SD rats were randomly divided into a normal group, a model group, an MTX group (1.5 mg·kg-1), and low-(40.8 g·kg-1) and high-(102.0 g·kg-1)combination groups (MTX+ Zhiwang decoction). The CIA model was induced in rats except for those in the normal group by tail injection of 50 μL of 2 g·L-1 type Ⅱ collagen on the 1st and 7th days. From the 8th day, the rats were treated correspondingly for 21 days. The body weight, toe thickness, and arthritis index(AI)were measured every seven days. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of inflammatory factors in the serum, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β(IL-1β). Tartrate-resistant acid phosphatase staining was used to detect the number of osteoclasts in the ankle joint. The expression of receptor activator of nuclear factor-κB ligand(RANKL) and osteoprotegerin(OPG) in ankle tissues was detected by immunohistochemistry(IHC), real-time polymerase chain reaction (Real-time PCR), and Western blot.ResultCompared with the normal group, the model group showed decreased body weight (P<0.01), increased toe thickness and AI score (P<0.01), up-regulated serum levels of TNF-α,IL-6, and IL-1β (P<0.01), elevated number of osteoclasts (P<0.01), increased mRNA and protein expression of RANKL (P<0.01), reduced mRNA and protein expression of OPG (P<0.05), and increased RANKL/OPG ratio (P<0.01). Compared with the model group,the high-dose combination group showed increased body weight,decreased toe thickness and AI score, reduced mRNA and protein expression of RANKL, up-regulated mRNA and protein expression of OPG, and declining RANKL/OPG ratio (P<0.05, P<0.01). The therapeutic effect was optimal in the high-dose combination group, followed by the low-dose combination group and the MTX group.ConclusionZhiwang decoction combined with MTX can improve bone destruction induced by rheumatoid arthritis, which may be achieved by inhibiting the RANKL/OPG pathway. This study provides an experimental basis for the treatment of bone destruction induced by rheumatoid arthritis with integrated traditional Chinese and western medicine.
Keywords:rheumatoid arthritis;Zhiwang decoction;receptor activator of nuclear factor-κB ligand(RANKL)/osteoprotegerin(OPG) pathway;bone destruction
Abstract:ObjectiveTo investigate the effect and mechanism of total flavones of Spatholobi Caulis (TFSC) against depression in rats.MethodThe fifty KM mice were randomly divided into the normal group and high-, medium-, and low-dose (1, 0.5, 0.25 g·kg-1) TFSC groups and gavaged with the corresponding drugs for 12 successive days. One hour after the last administration, the immobility time in forced swimming test and tail suspension test was recorded. The SD rats were randomly divided into the normal group, model group, fluoxetine (5 mg·kg-1) group, and high- and low-dose (1, 0.25 g·kg-1) TFSC groups. Following the exposure of rats to two different kinds of stimuli daily for inducing chronic unpredictable stress, they were administered with the corresponding drugs for 21 d. After the experiment, the levels of serum neurotransmitters and inflammatory factors in rats were detected by enzyme-linked immunosorbent assay (ELISA). The changes in hippocampal neurons of rats were observed by hematoxylin-eosin (HE) and Nissl staining. The mRNA expression levels of nuclear factor-κB (NF-κB) and tumor necrosis factor-α (TNF-α) in the hippocampus of rats were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expression levels of cAMP-response element binding protein (CREB), phosphorylated CREB (p-CREB), and brain-derived neurotrophic factor (BDNF) in hippocampal tissues by Western blot.ResultCompared with the normal group, TFSC significantly shortened the immobility time of mice in tail suspension and swimming tests (P<0.05). Compared with the normal group, the model group exhibited reduced sucrose intake and wilderness activity (P<0.01), decreased 5-HT, DA, NE (P<0.05, P<0.01), MAO, IL-6, TNF-α (P<0.05, P<0.01), damaged neurons, increased mRNA levels of TNF-α and NF-κB (P<0.01), and down-regulated BDNF and CREB protein expression (P<0.05). Compared with the model group, TFSC significantly enhanced sucrose intake and wilderness activity of rats (P<0.05), increased the serum 5-HT, DA and NE (P<0.05, P<0.01), and decreased the serum MAO, IL-6, and TNF-α (P<0.05, P<0.01) as well as NF-κB and TNF-α mRNA expression (P<0.01), up-regulated the protein expression levels of BDNF and CREB (P<0.01), and improved the pathological symptoms of hippocampus.ConclusionTFSC improved the hippocampal neurons of rats via CREB/BDNF signaling pathway and reduced depressive pathological damage, thus relieving depression.
Keywords:Spatholobi Caulis;depression;brain-derived neurotrophic factor (BDNF);cAMP-response element binding protein (CREB);neuroplasticity
Abstract:ObjectiveTo explore the mechanism of antidepressant effect of lily polysaccharide (LLP)and astragalus polysaccharide(APS).MethodSixty KM mice were randomly divided into blank group, model group, fluoxetine hydrochloride (8 mg·kg-1)group, LLP (0.2 g·kg-1)group, APS (0.2 g·kg-1)group and polysaccharide combination (LLP+APS,0.1 g·kg-1+0.1 g·kg-1)group, with 10 mice in each group. Except the blank group, the other groups were given chronic unpredictable mild stress (CUMS) induced mouse depression model. On the 29th day of modeling,fluoxetine hydrochloride group was given corresponding dose of fluoxetine hydrochloride, and polysaccharide groups were given corresponding drug. The depressive behavior of mice was evaluated by behavioral indexes such as body mass change, open field test. The morphological changes of hippocampal CA1 neurons were observed by Nissl staining. The contents of 5-hydroxytryptamine (5-HT), adrenocorticotropic hormone (ACTH), and corticosterone (CORT), in brain tissue and plasma were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression levels of related proteins in adenylate cyclase/cyclic adenylate phosphate/protein kinase A (AC/cAMP/PKA) signal pathway.ResultCompared with the blank group, mice in the model group gained weight slowly, the total distance, central distance and sugar water preference rate decreased significantly (P<0.01), the depressive behavior was significant, the hippocampal neurons were seriously damaged, the content of 5-HT decreased (P<0.01), the contents of ACTH and CORT increased significantly (P<0.01), adenylate cyclase 6(ADCY6), PKA and cAMP response element binding protein-1 (CREB-1) and brain-derived neurotrophic factor (BDNF) protein expression decreased significantly (P<0.01). Compared with the model group, depressive behavior of mice in LLP group, APS group and LLP+APS group was significantly improved (P<0.01). The antidepressant effect of LLP+APS was better than that of LLP and APS. Each administration group could alleviate the damage of hippocampal neurons in varying degrees, significantly increase the content of 5-HT in brain tissue (P<0.01), and reduce the levels of ACTH and CORT in plasma (P<0.05). The protein levels of ADCY6, PKA, CREB-1 and BDNF were significantly increased (P<0.05).ConclusionThe antidepressant effect of LLP+APS is significantly enhanced and has a synergistic effect. The mechanism may be closely related to affecting the content of neurotransmitters, inhibiting HPA axis activity and activating AC/cAMP/PKA signal transduction pathway.
Abstract:ObjectiveTo explore effect of Huanglian Jiedutang (HLJDT) on autophagy-related protein expression in septic mice with liver injury induced by cecal ligation and puncture (CLP).MethodSixty eight-week-old C57BL/6 mice were randomly divided into four groups, namely, the sham operation group, model group, and low- (1.44 g∙kg-1) and high-dose (2.88 g∙kg-1) HLJDT groups, with 15 in each group. The septic model was established by CLP after the last administration of HLJDT for three successive days. The survival rate of mice with 24 h was observed. The mice were sacrificed 12 h after operation for collecting the serum and liver tissue. The levels of serum interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA), and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum by biochemical method. The pathological changes in liver tissue were observed by hematoxylin-eosin (HE) staining, and the apoptosis index (AI) of hepatocytes by TdT-mediated dUTP-biotin nick end-labeling (TUNEL). The expression levels of protein high-mobility group box 1 protein (HMGB1), Beclin1, and microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ/Ⅰ in the liver tissue were assayed by Western blot.ResultCompared with the sham operation group, the model group showed reduced survival rate at 12 and 24 h, elevated IL-6, TNF-α, and IL-1β levels, enhanced AST and ALT activities (P<0.05), hepatocyte swelling, inflammatory cell infiltration, and apoptosis, and up-regulated HMGB1 (P<0.05), Beclin1, and LC3-Ⅱ/Ⅰ (P<0.05). Compared with the model group, each medication group exhibited increased survival rate at 12 and 24 h, lowered IL-6 and TNF-α levels, weakened AST and ALT activities (P<0.05), alleviated liver injury and apoptosis (P<0.05), down-regulated HMGB1 expression ( P<0.05), and up-regulated Beclin1 and LC3-Ⅱ/Ⅰ (P<0.05).ConclusionHLJDT alleviates the liver injury of septic mice possibly by inducing autophagy and inhibiting apoptosis.
Abstract:ObjectiveTo explore the differences in response to bakuchiol-induced hepatotoxicity between Institute of Cancer Research (ICR) mice and Kunming (KM) mice.MethodThe objective manifestations of bakuchiol-induced hepatotoxicity in mice were confirmed by acute and subacute toxicity animal experiments, and enrichment pathways of differential genes between normal ICR mice and KM mice were compared by transcriptomics. The real-time quantitative polymerase chain reaction (real-time qPCR) assay was used to verify the mRNA expression of key genes in the related pathways to confirm the species differences of bakuchiol-induced liver injury.ResultIn the subacute toxicity experiment, compared with the normal mice, the ICR mice showed increased serum content of alkaline phosphatase (ALP), and 5′-nucleotidase (5′-NT), without significant difference, and no manifest change was observed in KM mice. Pathological results showed that hepatocyte hypertrophy was the main pathological feature in ICR mice and hepatocyte steatosis in KM mice. In the acute toxicity experiment, KM mice showed erect hair, mental malaise, and near-death 3 days after administration. The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in KM mice (400 mg·kg-1) significantly increased(P<0.01), and the activity of total reactive oxygen species (SOD) in liver significantly decreased(P<0.01)compared with those in normal mice, while the serum content of 5′-NT and cholinesterase (CHE) in ICR mice (400 mg·kg-1) were significantly elevated (P<0.01). The liver/brain ratio in ICR mice increased by 20.34% and that in KM mice increased by 29.14% (P<0.01). The main pathological manifestation of the liver in ICR mice was hepatocyte hypertrophy, while those in KM mice were focal inflammation, hepatocyte hypertrophy, and hepatocyte steatosis. Kyoto Encyclopedia of Genes and Genomes(KEGG)and Reactome pathway enrichment analyses showed that the differential gene expression between ICR mice and KM mice was mainly involved in oxidative phosphorylation, bile secretion, bile acid and bile salts synthesis, and metabolism pathway. CYP7A1 was up-regulated in all groups with drug intervention (P<0.01) and MRP2 was reduced in all groups with drug intervention of KM mice (P<0.01) and elevated in all groups with drug intervention of ICR mice (P<0.01) compared with those in the normal group. The expression of BSEP was lowered in ICR mice with acute liver injury (400 mg·kg-1) (P<0.05). SHP1 was highly expressed in KM mice with acute liver injury (400 mg·kg-1). The expression of FXR was diminished in ICR mice with subacute liver injury (200 mg·kg-1) (P<0.01). SOD1, CAT, and NFR2 significantly decreased in KM mice with acute liver injury (400 mg·kg-1), and CAT dwindled in KM mice with subacute liver injury (200 mg·kg-1) (P<0.01). GSTA1 and GPX1 significantly increased in KM mice with acute liver injury (400 mg·kg-1) (P<0.01) and SOD1, CAT, NRF2, and GSTA1 significantly increased in ICR mice with subacute liver injury (200 mg·kg-1) (P<0.01). CAT and NRF2 significantly increased in ICR mice with acute liver injury (400 mg·kg-1) (P<0.01).ConclusionWith the increase in the dosage of bakuchiol, the liver injury induced by oxidative stress in KM mice was gradually aggravated, and ICR mice showed stronger antioxidant capacity. The comparison of responses to bakuchiol-induced hepatotoxicity between ICR mice and KM mice reveals that ICR mice are more suitable for the investigation of the mechanisms related to bile secretion and bile acid metabolism in the research on bakuchiol-induced hepatotoxicity in mice. KM mice are more prone to liver injury caused by oxidative stress.
Abstract:ObjectiveTo observe the long-term safety of Zhenwutang on blood indexes and histopathology in SD rats.MethodThe acute toxicity test was conducted to evaluate the acute toxicity of Zhenwutang toward SD rats. The SD rats were randomly assigned into 4 groups (low-, medium-, and high-dose Zhenwutang and control group), with 32 rats (16 males and 16 females) in each group. The rats were administrated once daily with Zhenwutang respectively at the doses of 16, 24, and 36 g·kg-1 (equal to 20, 30, 45 times of clinical dose) for 30 days, and those in the control group with purified water. After administration, the rats were observed for another 14 days. The body weight and food intake were measured once a week and the dosage was changed according to the body weight. General observation was carried out once a day during the experiment period. The electrocardiogram (ECG) was measured a day before administration (D-1), 4 h after administration (D0), on day 15 of administration (D14), and at the end of administration (D29) and the end of recovery period (D43). At the end of administration (D30) and the end of recovery period (D44), the blood assay and histopathological examinations were performed for 64 rats.ResultCompared with control group,some rats in zhenwutang high-dose group (D8-D35), medium-dose group (D13-D35) and low-dose group (D15-D33) showed depilation, mainly in the neck.Compared with the control group, rat body weight of D7 in Zhenwutang medium-dose and high-dose groups had significantly decreased,rat body weight of D21 and D29 in Zhenwutang low-dose group had decreased too.During D7-D29, body weight of rats in Zhenwutangn medium-dose and high-dose groups had differently decreased.Compared with control group, food intake in Zhenwutang low-dose, medium-dose and high-dose groups had significantly reduced in the first week. The food intake of male rats in Zhenwutang medium-dose and high-dose groups had significantly decreased in the first, second and fourth week .Compared with control group, the heart rate of female rats in Zhenwutang medium-dose and high-dose groups had increased. In the D14 and D29 days, the heart rate of male rats in Zhenwutang low-dose, medium-dose and high-dose groups had increased. The P wave time of male rats in Zhenwutang medium-dose group have prolonged. The QRS interval of female rats in Zhenwutang medium-dose and high-dose groups had prolonged. After 14 days of administration, the s-wave amplitude of Zhenwutang high-dose group had significantly decreased. After D14 and D29 days of administration, the s-wave amplitude of Zhenwutang in middle and high dose groups had significantly decreased.This study showed that intragastric administration of Zhenwutang for 30 days caused hair loss, weight loss, poor appetite, increased heart rate, and irregular heart rate of SD rats. It had little effect on the hematological indicators and did not cause obvious changes in pathological and blood indicators.ConclusionZhenwutang exerts positive inotropic effect, and 45 times of the clinical dose has no obvious toxic effect. However, attention should be paid to preventing the occurrence of ventricular premature beat.
Abstract:ObjectiveTo explore the clinical effect of modified Wuhutang in the treatment of children with acute asthma.MethodA total of 130 children with acute asthma were randomly divided into an observation group and a control group, with 65 cases in each group. The observation group was treated with modified Wuhutang and the control group was treated with procaterol hydrochloride for one week. The scores of primary symptoms (wheezing, cough, shortness of breath, and chest tightness) and secondary symptoms (mental status, runny nose, dry mouth, tongue texture, tongue coating, stool, etc.), lung functions, immunoglobulin E (IgE) expression, eosinophil (EOS) count, and serum inflammatory factors, including interleukin (IL)-5, IL-6, IL-8, and IL-1β in two groups before and after treatment were compared.ResultThe data of 126 children were statistically analyzed. As revealed by the results, compared with the conditions before treatment, the scores of primary symptoms and secondary symptoms, serum levels of IL, IgE expression, and EOS count were both reduced in two groups (P<0.05), lung functions were increased in the two groups(P<0.05). Compared with the control group after treatment, the observation group showed decreased scores of cough and secondary symptoms (P<0.05), and insignificant decrease in IL-1β (P<0.05). The improvement in lung functions, IgE expression, and EOS count in observation group was equivalent to that in control group.ConclusionModified Wuhutang for treatment of acute asthma in children (phlegm-heat obstructing lung syndrome) can significantly relieve the clinical symptoms, improve lung functions, and reduce IgE, IL-5, IL-6, IL-8, IL-1β expression levels and EOS count, and its overall clinical efficacy is superior or equivalent to that of tprocaterol hydrochloride.
Keywords:childhood asthma;acute exacerbation;modified Wuhutang;clinical research
Abstract:ObjectiveTo observe the clinical efficacy of modified Tuoli Xiaodusan (TLXDS) in adjuvant treatment of Helicobacter pylori (Hp)-positive peptic ulcer (PU) with cold-heat complex syndrome and explore its regulating effect on invasive/protective factors.MethodA total of 136 patients were randomly assigned into the control group (68 cases, including 4 cases missing, 3 cases eliminated, and 61 cases completed) and the TLXDS group (68 cases, including 4 cases missing, 1 case eliminated, and 63 cases completed). Both groups adopted the quadruple therapy of acid suppression and Hp eradication. The patients in the control group received Weinai'an capsules orally at 4 capsules/time and 3 times/day, and those in the TLXDS group took modified TLXDS orally at 1 dose/day. The administration of both groups lasted for 8 consecutive weeks and the follow-up lasted for 12 months. Electronic gastroscopy was carried out before and after treatment for evaluating the healing of ulcer, the score of mucosal morphology, and the maturity of regenerated mucosa. The Hp infection and the score of cold-heat complex syndrome were evaluated before and after treatment. The serum levels of gastrin (GAS), prostaglandin E2 (PGE2), pepsinogen (PG)-Ⅰ, PG-Ⅱ, epidermal growth factor (EGF), and trefoil factor 2 (TFF-2) were determined before and after therapy. The recurrence of Hp and PU was recorded, and the drug safety was evaluated.ResultAfter treatment, the mucosal morphology score and the traditional Chinese medicine (TCM) syndrome score in the TLXDS group were lower than those in the control group (P<0.01). The levels of GAS, PG-Ⅰ, and PG-Ⅱ in the TLXDS group were lower than those in the control group (P<0.01), whereas those of PGE2, EGF, and TFF-2 showed an opposite trend (P<0.01). After treatment, the Hp eradication rate in the TLXDS group was 95.24% (60/63), higher than that (83.61%, 51/61) in the control group (χ2=4.467, P<0.05). The total effective rate of TCM syndromes in the TLXDS group was 98.41% (62/63), higher than that (81.97%, 50/61) in the control group (χ2=9.589, P<0.01). The total effective rate of the TLXDS group under gastroscopy was 98.41% (62/63), higher than that (86.89%, 53/61) in the control group (χ2=4.525, P<0.05). The excellent and good rate of regenerated mucosal maturity in the TLXDS group was 92.06% (58/63), also higher than that (73.77%, 45/61) in the control group (χ2=7.372, P<0.01). After 12 months of follow-up, the TLXDS group had lower PU recurrence rate [19.05% (12/63) vs 37.70% (23/61), χ2=5.325, P<0.05] and lower Hp recurrence rate [15.00% (9/60) vs 33.33% (17/51), χ2=5.165, P<0.05) than the control group. No adverse reactions related to TLXDS were detected.ConclusionModified TLXDS-assisted quadruple therapy demonstrates significant short-term clinical efficacy and high Hp eradication rate for Hp-positive PU (cold-heat complex syndrome) patients. Moreover, it can adjust the levels of invasive/protective factors to facilitate ulcer healing and reduce the recurrence rates of Hp and PU in a long term, with good clinical safety.
Abstract:ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout (ApoE-/-) mice, and explore its mechanism of treating atherosclerosis by regulating intestinal flora.MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group, rosuvastatin group (10 mg·kg-1), high-, low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis (75, 25 mg·kg-1), with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration, blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin (HE) staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora.ResultCompared with the blank group, the plaque area of the model group was significantly increased with inflammatory infiltration, the contents of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), inflammatory factors and inducible nitric oxide synthase (iNOS) were increased, while the content of high-density lipoprotein cholesterol (HDL-C) was decreased. Compared with the model group, rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque, reduce the contents of TG, TC, LDL-C, inflammatory factors and iNOS, and increase the content of HDL-C. Compared with the blank group, the relative abundances of Firmicutes and Proteobacteria in the model group increased, while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated, and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group, ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria.ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids, which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors, improve antioxidant capacity and repair the disorder of intestinal flora structure.
Keywords:Acanthopanacis Senticosi Radix et Rhizoma seu Caulis;ethyl acetate extract;intestinal flora;apolipoprotein E gene knockout (ApoE-/-) mice;atherosclerosis;inflammatory factor;antioxidant activity;vascular endothelial factor
Abstract:ObjectiveTo establish a high performance liquid chromatography (HPLC) for simultaneous determination of baicalin magnesium and baicalein in rat plasma and tissues, and to investigate the effect of acute liver injury on pharmacokinetics and tissue distribution of baicalin magnesium in rats.MethodAcute liver injury rat model was induced by carbon tetrachloride (CCl4). Normal rats and acute liver injury model rats were given an equal dose (287.31 mg·kg-1) of baicalin magnesium aqueous solution by intragastric administration, the orbital blood was collected at different time points, and HPLC was used to simultaneously determine the concentrations of baicalin magnesium and baicalein in rat plasma at each time point, the concentration-time curves were drawn, the pharmacokinetic parameters were calculated with DAS 3.0, and SPSS 23.0 was used for statistical analysis. After oral administration of baicalin magnesium aqueous solution, HPLC was used to simultaneously determine the contents of baicalin magnesium and baicalein in rat liver, lung, kidney, stomach, brain and small intestine at different time points, the mobile phase was 0.1% phosphoric acid aqueous solution-methanol, and the detection wavelength was 278 nm.ResultIn the acute liver injury model group, the peak concentration (Cmax) of baicalin magnesium was 0.58 times that of the normal group, the area under concentration-time curve (AUC0-t) was 0.5 times that of the normal group (P<0.05), the apparent volume of distribution (Vd) was 2.3 times that of the normal group (P<0.05), and baicalein is almost undetectable in plasma. The content of baicalin magnesium in liver, stomach and brain of the acute liver injury model group was higher than that of the normal group at each time point, while the content of baicalin magnesium in the samples of lung at 8 h, kidney at 8 h and 12 h, and small intestine at 0.333 h was lower than that of the normal group. The content of baicalein in lung, stomach and small intestine of the model group was higher than that of the normal group at each time point, while the content of baicalein in the tissue samples of liver at 6, 8 h and kidney at 0.333, 4, 6 h was lower than that in the normal group, and baicalein could hardly be detected in the brain.ConclusionAfter intragastric administration of the same dose of baicalin magnesium aqueous solution, acute liver injury induced by CCl4 can affect the pharmacokinetics and tissue distribution characteristics of baicalin magnesium in rats, and there is biotransformation of baicalin magnesium and baicalein in liver, lung, kidney, stomach and small intestine.
Abstract:ObjectiveTo explore the effects of three kinds of microbial fertilizers on the growth, yield, quality, and cadmium (Cd) accumulation of Chuanxiong Rhizoma (CX).MethodTaking CX seeds as materials, field experiments were carried out in the main producing areas, Pengzhou and Meishan. The samples were collected during the harvesting period, and the agronomic characters and yield were determined. The contents of extract, volatile oil, and ferulic acid were analyzed by the collection method of Chinese Pharmacopoeia (2020 edition). The content of Cd was determined by inductively coupled plasma-mass spectrometry (ICP-MS). Data were processed by difference significance analysis, correlation analysis and cluster analysis.ResultThree kinds of microbial fertilizers with appropriate concentrations could promote the growth of CX. In terms of yield and quality, the treatment of Jinwuzong (1.50 ton/hm2, 1 ton=1 000 kg, the same below) and Cuijingyuan (1.5 L·hm-2) could increase the yield of medicinal materials by 0.92%-46.34%, while Cuijingyuan (1.8 L·hm-2) and Shenchu (15, 30 kg·hm-2) could increase the water-soluble extract of CX by 0.06%-18.79%, of which Cuijingyuan (1.8 L·hm-2) was significantly increased (P<0.01). The alcohol-soluble extract of CX treated with Jinwuzong (0.75, 1.50, 2.25 ton/hm2), Shenchu (15, 45 kg·hm-2), and Cuijingyuan (1.2 L·hm-2) decreased significantly by 3.51%-22.94% (P<0.01). The content of ferulic acid in CX treated with Jinwuzong (1.50 ton/hm2) and Shenchu (30 kg·hm-2) decreased by 2.14%-30.56%. Three kinds of microbial fertilizers had little effect on the content of volatile oil in CX. In the aspect of Cd enrichment, the concentration of Cd in rhizosphere soil of CX was increased by 11.33%-76.36% (P<0.01) after the treatment of Jinwuzong (0.75, 1.50, 2.25 ton/hm2), Shenchu (15, 30, 45 kg·hm-2) and Cuijingyuan (1.2 L·hm-2). However, the Cd enrichment coefficient of CX reduced by 2.58%-48.38%, the Cd content and Cd accumulation of CX decreased respectively by 9.54%-25.96% and 9.34%-18.88% via Jinwuzong (0.75 ton/hm2) and Cuijingyuan (1.8 L·hm-2).ConclusionThree kinds of microbial fertilizers have a certain positive effect on the growth, substance accumulation, and reduction of Cd content in medicinal parts of CX, and the changes of each index are affected by the producing area and treatment method. Based on the comprehensive analysis of various indicators, Jinwuzong (0.75, 1.50 ton/hm2) can better adapt to the rhizosphere soil micro-ecological environment of CX, it can effectively reduce the content of Cd on the premise of guaranteeing the yield and quality of CX.
Keywords:Chuanxiong Rhizoma;microbial fertilizer;growth and development;yield;quality;heavy metals;cadmium (Cd) enrichment
Abstract:ObjectiveTo screen the appropriate reference genes for real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR)analysis of the Andrographis paniculata under methyl jasmonate(MeJA)and various abiotic stresses.MethodThe actin 1(ACT1),actin 2(ACT2),elongation factor(EF-1α),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),tubulin(TUB),polyubiquitin(UBQ), and 18S rRNA(18S)gene were selected as candidate reference genes based on the RNA-seq data of high temperature,drought, UV, and MeJA. The expression of seven candidate reference genes in the A. paniculata leaves was assessed by Real-time PCR,and the stability was analyzed by geNorm,NormFinder,BestKeeper, and Refinder.ResultThe results of stability evaluated by geNorm,NormFinder, and BestKeeper were not the same due to different indicators. As analyzed by Refinder, for the stability of the expression, the genes were ranked as UBQ>18S>EF-1α>ACT2>ACT1>GAPDH>TUB under high temperature stress, ACT1>UBQ>EF-1α>18S>ACT2>GAPDH>TUB under drought stress, EF-1α>TUB>ACT2>UBQ>18S>GAPDH>ACT1 under UV stress, and ACT1>EF-1α>UBQ>ACT2>18S>TUB>GAPDH under MeJA stress. Among them,18S gene was not suitable as an internal reference gene duo to its high expressive abundance. This study also verified the relative expression level of andrographolide synthesis-related gene hydroxy-methylglutaryl-CoA synthase (HMGS) in the four stresses on the basis of transcriptome data,and found that the Real-time PCR results of appropriate internal reference genes were accurate and reliable.ConclusionUBQ-ACT1-UBQ,EF-1α-TUB,and ACT1-EF-1α were the suitable combinations under stresses of high temperature,drought,UV, and MeJA. This study is expected to provide references for the research on function regulation and expression of genes in A. paniculata under high temperature,drought,UV, and MeJA stresses.
Abstract:ObjectiveTo study the in vitro kinetics of Jiaojiang cataplasms and evaluate its pharmacodynamics, so as to provide a feasible basis for the development of this preparation.MethodThe improved Franz diffusion cell was used for the in vitro release in semipermeable membrane and transdermal absorption in in vitro mouse skins. The contents of hydroxy-α-sanshool, 6-gingerol, ginsenoside Rb1 were determined by high performance liquid chromatography (HPLC), to evaluate the in vitro release and transdermal absorption of Jiaojiang cataplasms. The mobile phase of 6-gingerol and hydroxy-α-sanshool was water-acetonitrile-methanol (2∶1∶1) with the detection wavelength of 280 nm. The mobile phase of ginsenoside Rb1 was acetonitrile-0.1% phosphoric acid aqueous solution (31∶69) with the detection wavelength of 203 nm. A mouse intestinal paralysis model was established, and mice were randomly divided into five groups, namely sham operation group, model group, domperidone group (3.9 mg·kg-1) and high- and low-dose groups of Jiaojiang cataplasms (6.2, 3.1 g·kg-1, measured by crude drug dosage), to observe the effect of this preparation on gastrointestinal propulsion function.ResultAverage release rates of hydroxy-α-sanshool, 6-gingerol and ginsenoside Rb1 at 24 h were 16.41, 4.23, 4.15 μg∙cm-2∙h-1, the average transdermal rates of them at 24 h were 2.31, 0.64, 0.29 μg∙cm-2∙h-1, their skin retention values were 19.56, 3.59, 1.61 μg, respectively. According to the Ritger-Peppas equation, the release of hydroxy-α-sanshool, 6-gingerol, ginsenoside Rb1 was non-Fick diffusion. The high-dose group of Jiaojiang cataplasms could improve intestinal function of model mice after small intestinal friction injury, and promote intestinal peristalsis and small intestinal propulsion rate (P<0.05).ConclusionJiaojiang cataplasms has in vitro release and transdermal properties, the in vitro release conforms to Higuchi equation, and transdermal absorption behavior conforms to zero-order kinetic equation, which can improve the postoperative function of the small intestine and the propulsion function of small intestine. It preliminarily indicates that the preparation has certain clinical development value.
Abstract:ObjectiveTo investigate the effects of micro-fertilizer containing rare earth of different types and concentrations on the growth,yield and quality of Angelica sinensis.MethodOn the basis of the single-factor randomized block design, the growth and index components of Angelica sinensis were determined with rare earth-containing nitrate and chloride micro-fertilizers of different concentrations as foliar fertilizers.ResultSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer could increase the economic yield of A. sinensis, with the fresh yield per mu (1 mu≈667 m2) reaching 855.4 kg and the dry yield per mu 350.7 kg,which increased by 15.16% and 28.70% respectively compared with that in the control group CK1. Spraying 1.2 g·mL-1 rare earth-containing nitrate micro-fertilizer could promote the growth and development of A. sinensis and significantly increase the content of index components, with the plant height reaching 93.05 cm,the stem diameter 15.60 mm,the root diameter 16.10 mm,the main root length 36.5 cm,and the number of leaves 11.25 pieces per plant, which increased by 32.76%,31.98%,41.98%,53.36%,and 45.16%, respectively, compared with those in the control group CK2. Besides, the content of ferulic acid,volatile oil,ligustilide, and extract was 0.96%,0.41%,0.30% and 48.76%,respectively,which increased by 12.94%,17.14%,11.11%, and 12.07%,respectively,compared with that in the control group CK2.ConclusionSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer and 1.2 g·mL-1 rare earth-containing nitratemicro-fertilizer can promote the growth and development of A. sinensis,improve the medicinal properties,and increase yield and quality. Rare earth-containing micro-fertilizers can be applied in the standardization of A. sinensis cultivation, which can change the production status of A. sinensis that depends on chemical fertilizers and single fertilization, and promote the green, organic and ecological cultivation of A. sinensis.
Keywords:Angelica sinensis;micro-fertilizer containing rare earth;growth and development;yield and quality
Abstract:ObjectiveTo explore the mechanism of herbal pair Astragali Radix-Puerariae Lobatae Radix (AR-PLR) against type 2 diabetes mellitus (T2DM) based on network pharmacology and experimental verification.MethodThe active ingredients and targets of AR and PLR were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The related targets of T2DM were retrieved from disease databases and the common targets of drugs and diseases were extracted. The protein-protein interaction (PPI) network was analyzed and constructed by STRING and the network topology of key targets was analyzed by Cytoscape 3.7.1. Then gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analyses of core targets were carried out by DAVID to explore its possible molecular mechanism. The T2DM model was induced in rats by the high-fat diet combined with tail intravenous injection of streptozocin. The rats were divided into a normal group,a model group,a metformin group,and high-,medium- and low-dose AR-PLR groups. After four weeks of intragastric administration,the serum levels of fasting blood sugar (FBS),fasting insulin(FINS),aspartate aminotransferase(AST),alanine aminotransferase(ALT),triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterin(LDL-C),high-density lipoprotein cholesterin (HDL-C),interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) of rats in each group were measured. The protein expression of insulin receptor substrate-2(IRS-2),phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt), and forkhead box transcription factor O1(FoxO1) in rat liver was detected by Western blot.ResultA total of 131 core targets of AR-PLR in the treatment of T2DM were screened out by network pharmacology, where Akt1,mitogen-activated protein kinase 1 (MAPK1),TNF-α,and IL-6 were critical. As revealed by KEGG enrichment analysis, AR-PLR exerted the hypoglycemic effect mainly through the PI3K/Akt,TNF, and FoxO signaling pathways. Compared with the model group,the high- and medium-dose AR-PLR groups showed reduced FBS and FINS levels and increased glycogen level (P<0.05,P<0.01),all the AR-PLR groups showed decreased levels of AST,ALT,TG, and LDL-C (P<0.05,P<0.01), the high- and low-dose AR-PLR groups showed decreased TC levels (P<0.05). Compared with the model group, the high- and medium-dose AR-PLR groups showed reduced levels of IL-6 and TNF-α(P<0.05,P<0.01), and the high-dose AR-PLR group showed increased expression of IRS-2, Akt, p-Akt, PI3K, and p-PI3K, and decreased expression of FoxO1 protein(P<0.05).ConclusionAR-PLR has the characteristics of multi-component,Multi-target and multi-pathway in the treatment of T2DM. This herbal pair may regulate the PI3K/Akt/FoxO1 signaling pathway through IL-6, TNF-α, and other targets to affect insulin resistance, glycogen synthesis, gluconeogenesis, glucose transport, inflammation, immune response, and other processes, thereby treating T2DM.
Abstract:ObjectiveTo investigate the potential mechanism of Xiao Chaihutang (XCHT) in the treatment of Alzheimer's disease (AD) based on network pharmacology and bioinformatics.MethodThe active components of XCHT and corresponding targets were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the differentially expressed genes related to AD were searched from Gene Expression Omnibus (GEO). Thereby, the common targets of XCHT and AD were yielded, followed by Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the common targets. The component-target network and protein–protein interaction (PPI) network were constructed. Furthermore, amyloid β-protein (Aβ)1-40 was used to induce AD in PC12 cells and then the AD cells were intervened with XCHT. Afterward, cell viability was detected by Cell Counting Kit-8 (CCK-8) assay and cell morphology was observed based on 4',6-diamidino-2-phenylindole (DAPI) staining. Cell membrane potential was determined and apoptosis was detected by flow cytometry, and cellular immunofluorescence detects the expression of B-cell lymphoma-2 (Bcl-2)/Bcl-2-related X protein (Bax). Moreover, immunofluorescence assay was performed.ResultA total of 190 active components and 41 anti-AD targets of XCHT were screened out. The key components included mairin, quercetin, berberine, protoporphyrin, 24-ethylcholest-4-en-3-one, and β-D-ribofuranoside, and the core targets were sigma non-opioid intracellular receptor 1 (SIGMAR1), checkpoint kinase 1 (CHEK1), protein tyrosine phosphatase non-receptor type 6 (PTPN6), protein kinase C(PRKCH), inhibitor of nuclear factor kappa B kinase subunit beta (IKBKB), cathepsin D (CTSD), cysteine aspartate protease-3 (Caspase-3), Bax, and Bcl-2-like protein 1 (Bcl-2L1). The anti-AD targets of XCHT were involved in 302 GO terms (P < 0.05), particularly the regulation of neuronal cell apoptosis, and 73 KEGG pathways (P<0.05). The major pathways and biological processes included the apoptosis pathway, virus infection pathway, lipid and atherosclerosis pathway, and cancer-related pathways. In the in vitro experiment, the model group demonstrated the decrease in cell survival rate (P<0.05), increase in apoptosis rate (P<0.05), and down-regulation of mitochondrial membrane potential and Bcl-2/Bax ratio compared with the blank control. Compared with the model group, XCFT group showed the increase in cell survival rate (P<0.05), decrease in apoptosis rate (P<0.05), and up-regulation of mitochondrial membrane potential and Bcl-2/Bax ratio.ConclusionBased on network pharmacology, this study reveals the multi-component, multi-target, and multi-pathway characteristics of XCHT in the treatment of AD, laying a foundation for further research on the material basis and mechanism of this prescription.
Abstract:ObjectiveTo explore the underlying mechanism of bile acids and metabolites as well as the key metabolic pathways and important endogenous targets in prehypertension.MethodThe metabolic mechanism of prehypertension was explored with non-targeted metabolomics combined with network analysis. The serum metabolomics of patients with prehypertension was analyzed by ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry. The relevant biological functions and signal targets were predicted and generated by network analysis. Finally,the predicted targets of this important pathway were verified by in vitro experiments,and the relevant information was verified by enzyme-linked immunosorbent assay (ELISA) and Western blot.ResultAs revealed by non-targeted metabolomics,there were 64 potential biomarkers and 13 metabolic pathways in the normal group,the prehypertension group, and the hypertension group. The results of network analysis and biological verification showed that the occurrence of prehypertension was related to vascular inflammation caused by the abnormal metabolism of bile acids and aromatic amino acids. Bile acid metabolism plays an important role in the occurrence and development of prehypertension by regulating the vascular inflammatory response. Amino acid N-acyltransferase,myeloperoxidase, and bile acid downstream receptor TGR5 are critical in the changes of the metabolic network.ConclusionIn prehypertension,bile acids are presumedly involved in regulating vascular inflammation, resulting in damage to blood vessels in prehypertension.
Abstract:ObjectiveBased on the medical cases of Qi and blood co-treatment of traditional Chinese medicine(TCM) masters, to discover the syndrome and treatment rules and medication experience of Qi and blood co-treatment through data mining.MethodFrom December 1999 to November 2020, the Qi and blood treatment cases of TCM masters were retrieved from China National Knowledge Infrastructure (CNKI). Frequency statistics, association rules, cluster analysis and other methods were used for data mining.ResultThe analysis of 591 medical cases of 57 national medical master found that blood stasis, Qi deficiency, Qi stagnation, blood deficiency and phlegm were the most common syndromes. The tongue was reddish, pale or dark, the moss was white or thin, and the pulse was thin, stringy, heavy and slippery. In the treatment of Qi and blood, the disease in the early stage is mostly in Qi and blood itself. At this time, the emphasis should be on regulating Qi and blood, or tonic or attack or both. At the same time, attention should be paid to invigorating the spleen, soothing the liver and tonifying the kidney. Core drugs include Danggui Buxuetang, Buyang Huanwutang, Huangqi Guizhi Wuwutang, Taohong Siwutang, Si Junzitang, Linggui Zhugantang, Xiaoyaosan, Danggui Shaoyaosan and other chemical cut.ConclusionWhen treating Qi and blood together, Chinese medical masters attach great importance to the relationship between Qi and blood and the development stage of diseases, and emphasize the precision and dynamic differentiation of treatment. Their theories and experience of diagnosis and treatment are worthy of clinical application and promotion.
Keywords:traditional Chinese medicine master;Qi and blood treatment;law of syndrome differentiation and treatment;clinical medical record;data mining
Abstract:ObjectiveTo systematically evaluate the clinical effectiveness and safety of Chinese medicinal injection (CMI) in the treatment of unstable angina pectoris (UAP).MethodEight databases, i.e., China National Knowledge Infrastructure (CNKI),VIP,Wanfang Data,CBM,PubMed,EMBASE,The Cochrane Library,and Web of Science were searched for randomized controlled trials (RCT) of conventional treatment combined with CMI (treatment group) versus conventional treatment (CT)(control group)in the treatment of UAP published from database inception to March 31th 2021. Stata 16.0 was used for network Meta-analysis.ResultThirty-nine RCT involving 3 407 patients were included. As revealed by the results of network Meta-analysis, in terms of the total effective rate in angina pectoris improvement, the therapeutic protocols were ranked as Tanreqing injection(TRQI)+CT>Xiangdan injection(XDI)+CT>Ciwujia injection(CWJI)+CT=Shengmai injection(SMI)+CT>Xuesaitong injection(XSTS)+CT>Breviscapine injection(BI)+CT>Shuxuetong injection(SXTI)+CT>Kudiezi injection(KDZI)+CT>Shuxuening injection(SXNI)+CT>Danshen injection (DSI)+CT>Guanxinning injection(GXNI)+CT>Dengzhanxixin injection(DZXXI)+CT>Xueshuantong injection(XSTI)+CT>Gualoupi injection(GLPI)+CT>CT;for the total effective rate in ECG improvement, SXTI+CT>XDI+CT>TRQI+CT>CWJI+CT>XSTI+CT>BI+CT>XSTI+CT>SXNI+CT>GXNI+CT>KDZI+CT>DZXXI+CT>GLPI+CT>CT>SMI+CT;for the adverse reactions, DZXXI+CT>XDI+CT>DSI+CT>BI+CT>SMI+CT>SXNI+CT>CT>GLPI+CT>GXNI+CT>SXTI+CT>KDZI+CT>CWJI+CT;for the reduction of fibrinogen (FIB), BI+CT>SXTI+CT>XSTI+CT>CT>KDZI+CT;for the reduction of C-reactive protein (CRP), DSI+CT>DZXXI+CT>XSTI+CT>CT;for the reduction of high-sensitivity C-reactive protein (hs-CRP), SXNI+CT>KDZI+CT>SXTI+CT>DZXXI+CT>GLPI+CT>TRQI+CT>XSTI+CT>CT. The results of subgroup analyses were consistent with those of the overall Meta-analysis.ConclusionCMI combined with CT can improve angina pectoris and ECG,reduce adverse reactions,and also improve FIB,CRP,and hs-CRP to varying degrees. However,due to the differences in the quality and quantity of CMIs in RCTs,clinical application should be performed based on the specific conditions.
Abstract:To give full play to the therapeutic advantages of traditional Chinese medicine (TCM) in sepsis, clarify the entry point of integrated TCM and western medicine, further standardize the clinical treatment of TCM, develop a recognized and integrated treatment protocol of TCM and western medicine, and improve the clinical efficacy on sepsis,the Chinese Association of Chinese Medicine organized TCM and western medicine experts specialized in sepsis treatment to conduct in-depth discussions on the advantages of TCM and integrated TCM and western medicine in the treatment of sepsis based on the TCM etiology and pathogenesis of sepsis, a representative acute and critical disease. They emphasized the pathogenesis characteristics of asthenia of healthy Qi and sthenia of pathogenic factors and summarized the roles of Chinese medicine in correcting the imbalance of inflammatory response, improving blood coagulation dysfunction, and relieving organ damage. Furthermore, they proposed the treatment protocol with integrated TCM and western medicine, which is expected to provide references for actual clinical treatment and scientific research.
Keywords:sepsis;diseases responding specifically to traditional Chinese medicine;expert consensus
Abstract:Inflammation is the key pathogenic feature of heart failure (HF), and its overexpression can cause myocardial hypertrophy, apoptosis and fibrosis, aggravating the process of HF. Macrophages are important immune cells in human body with high heterogeneity, which involve in inflammation response and maintain heart homeostasis. Macrophage polarization is a dynamic process. Under the stimulation of different microenvironment, it can polarize two subsets, including classically activated M1 type and alternatively activated M2 type, which are antagonistic to each other. When macrophages polarize into the pro-inflammatory phenotype (M1), the inflammatory response is initiated, the anti-inflammatory phenotype (M2) plays a role in inhibiting inflammation and repairing tissue. At the same time, in different stages of development of HF, M1 and M2 macrophages can be transformed into each other, which is similar to the connotation of Yin-yang restriction, balance and transformation of traditional Chinese medicine (TCM) theory. Based on this, this paper intends to clarify the relationship between M1 and M2 macrophages by Yin-yang theory, proposing that the clinical prevention and treatment of HF should pay attention to regulating micro and macro inflammatory responses, regulating macrophage polarization, and achieving the balance between anti-inflammatory and pro-inflammation, which is consistent with the balance of Yin-yang in TCM theory. It can provide a new target and direction for TCM treatment of HF.
Keywords:heart failure;Yin-yang theory;macrophage polarization;inflammation response;traditional Chinese medicine;microenvironment;cardiac homeostasis
Abstract:In the era of artificial intelligence based on big data, data acquisition, storage and processing are more convenient, which provides a guarantee for accelerating the development of traditional Chinese medicine (TCM), but it has not yet achieved organic integration with TCM theory. Based on preliminary research on the supramolecular "Qi chromatography" theory of TCM, combined with the current development trend of artificial intelligence, this paper analyzed the biological intelligence attribute of the function of TCM supramolecular "imprinting template", in order to provide reference for the development of TCM drug innovation. Both the human body and Chinese materia medica are giant complex supramolecular bodies evolved from natural organisms. According to the "imprinting template", the "social molecules" are controlled step by step to form the meridians and viscera. The interaction produces the original theory of TCM, in which the self-recognition, self-assembly, self-organization and self-replication of the "imprinting template" reflect the "intelligence" function attributes:the human body uses the "imprinting template" to self-identify and sense the ingredients of TCM, and store the memory information database in the meridian and collateral organs in the form of "imprinting template", and then pass the "imprinting template". The comparison, analysis, and judgment of imprinting templates guide the self-assembly, self-organization and self-replication among "molecular society", synthesize biological machines, produce biological functions, repair or strengthen biological supramolecular bodies, and present the most basic "intelligence" attribute. This suggests that the theory of theory-method-prescription-medicine of TCM is the weak embodiment of biological "intelligence", while the human brain function is the strong embodiment of biological "intelligence". Since the intelligent function of supramolecular "imprinting template" runs through the natural world, artificial intelligence that can characterize the strong "intelligence" form of the human brain will also be integrated into all aspects of the natural world, suggesting the development direction of "intelligence" functionalization of drug innovation mode.
Keywords:basic theory of traditional Chinese medicine;supramolecular "Qi chromatography" theory;Chinese materia medica;imprinting template;artificial intelligence;big data era
Abstract:With the population aging, the morbidity and mortality of cancer patients continue to rise. At present, the treatment methods for tumors include surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy. However, most chemotherapeutic drugs can cause severe side effects and drug resistance. Therefore, as an alternative therapy, traditional Chinese medicine (TCM) treatment can effectively relieve the clinical symptoms of tumor patients, improve the quality of life, inhibit or stabilize the development of tumors, and prolong the survival period of patients. Due to the good safety of Chinese medicine, its potential anti-cancer activity has attracted increasing attention. Ganoderma lucidum, a treasure of Chinese medicinal material, is a medicinal fungus with a history of more than 2 000 years in China. So far, many studies have proposed the anti-cancer properties of G. lucidum. G. lucidum has extensive pharmacological activities, such as anti-tumor, anti-atherosclerosis, and anti-aging. It can also regulate immunity, protect the liver and the heart, and reduce blood glucose and lipid. The chemical composition of G. lucidum is complex. At present, it is proved to contain polysaccharides, triterpenoids, alkaloids, nucleosides, amino acids, and various trace elements. The anti-tumor mechanisms of polysaccharides and triterpenoids in G. lucidum are mainly achieved by apoptosis induction, immune regulation, anti-angiogenesis, and induction of cell cycle arrest. Currently, it has been widely used in the adjuvant treatment of complex tumors such as lung cancer, liver cancer, cervical cancer, prostate cancer, breast cancer, and colon cancer. The present study reviewed the bioactivities and mechanisms of triterpenoids and polysaccharides in G. lucidum in recent years and highlighted the anti-tumor effects and mechanisms to provide references for the further development and utilization of G. lucidum.
Abstract:Prunellae Spica is the dry ear of the labiaceae plant Prunella vulgaris, which is a traditional medicine and food plant with many functions. Prunellae Spica can clear liver-fire, improve eyesight, disperse knot detumescence. It owns hot and bitter flavors and cold property. It goes to the liver, gallbladder meridian, and is a kind of commonly-used antifebric. Prunellae Spica has been used in the treatment of mammary gland diseases since ancient times.The mammary abscess, mammary nodules, mammary carcinoma of traditional Chinese medicine all belong to breast disease, and the liver meridian is most closely related to these diseases. With the development of social life, breast disease has gradually become the most primary health problem for women. Modern pharmacological studies show that Prunellae Spica contains terpenoids, phenolic acids, flavonoids and other biological active components, which have anti-inflammatory, antibacterial, hormone regulation, anti-tumor and other effects. Prunellae Spica inhibits the p38 mitogen-activated protein kinase (p38 MAPK)/nuclear factor kappa-B (NF-κB) pathway to play an anti-mastitis role, interferes with the effects of estrogen receptors or regulates lipid levels to treat breast hyperplasia, and treats breast cancer through promoting the apoptosis of breast cancer cells, inhibiting the migration of breast cancer cells, regulating the division of breast cancer cells and other ways. While referring to the relevant literature, it was found that Prunellae Spica often exerted pharmacological effects through multi-channels and multi-target regulation, but most of the studies did not specify the specific target of its effect, which needs further study. In this review, the effects and mechanisms of Prunellae Spica in the treatment of various breast diseases were summarized, so as to provide a reference for further research on the wider clinical therapeutic effects of Prunella subtilis and its therapeutic effects on breast diseases.
Keywords:Prunellae Spica;mammitis;hyperplasia of mammary glands;mammary cancer;pharmacological action
Abstract:Diabetic peripheral neuropathy is a common complication of diabetes, and its pathogenesis is complex. Its high morbidity can result in disability, teratogenesis, and death in diabetic patients. At present, the pathogenesis of diabetic peripheral neuropathy has not been clearly elucidated, which may be related to oxidative stress, inflammatory response, microcirculation dysfunction, metabolic abnormalities, etc. Recent studies have found that apoptosis plays an important role in the pathogenesis of diabetic peripheral neuropathy. The three pathways, i.e., mitochondrial pathway, death receptor pathway, and endoplasmic reticulum pathway, jointly regulate the cell apoptosis in the body. Traditional Chinese medicine, with definite efficacies in the treatment of diabetic peripheral neuropathy, is advantageous in overall regulation and multi-target and multi-pathway treatment. As reported, the active ingredients in Chinese medicine and Chinese medicinal compounds can alleviate diabetic peripheral neuropathy by regulating apoptosis signaling pathways. Furthermore, apoptosis pathways are expected to be potential targets for new drugs against diabetic peripheral neuropathy following oxidative stress. Therefore, this paper, taking apoptosis as the entry point, reviewed the research progress on TCM intervention in diabetic peripheral neuropathy in recent years to provide references for the clinical prevention and treatment of diabetic peripheral neuropathy and the development of new drugs.
Keywords:diabetic peripheral neuropathy;traditional Chinese medicine;cell apoptosis
Abstract:Atopic dermatitis (AD) is a chronic, recurrent and inflammatory skin disease. Modern medical research suggests that AD is related to immune function, genes, skin barrier and other factors, while the specific etiology and pathogenesis remains unclear. The disease has a long course and is prone to reoccur, which seriously affects people’s production and life. Steroids, antihistamines and immunosuppressants are commonly used western medicines for the treatment of AD, which, however, will cause adverse reactions after long-term application. Traditional Chinese medicine (TCM) has a long history, good therapeutic effect and rich clinical experience in the prevention and treatment of AD, and the research on the treatment of AD with TCM has been intensifying. Centering on the theory of TCM, we systematically summarized the research progress related to AD, discussed the etiology and pathogenesis of AD, and summarized the TCM syndrome differentiation of AD from the aspects of eight principles, etiology, Qi-blood-body fluid, zang-fu organs, six meridians, defense-Qi-nutrient-blood and triple energizer. According to the etiology and pathogenesis of AD, we proposed the therapeutic regimens corresponding to the type and stage of the disease. Considering the research progress achieved in the recent years in the prevention and treatment of AD by TCM and the modern pharmacological research on Chinese medicinal materials, we reviewed the classic famous prescriptions, self-made prescriptions and Chinese patent medicines, and expounded the mechanisms of single Chinese medicinal materials in the treatment of AD at the molecular level. The TCM external therapies such as wet compress, medicated bath, gel and ointment are safe and effective. Acupuncture and moxibustion play a role in the prevention and treatment of AD, which is worthy of promotion in clinical practice, and the syndrome differentiation methods of Qi-blood-body fluid and triple energizer are novel in the treatment of this disease. TCM, characterized by diverse therapeutic methods and good clinical efficacy, is worthy of promotion in the treatment of AD, which will contribute to the development of TCM in China.
Keywords:atopic dermatitis (AD);traditional Chinese medicine;syndrome differentiation and treatment;skin disease
Abstract:Postmenopausal osteoporosis (PMOP) is a systemic disease characterized by increased bone fragility caused by insufficient estrogen secretion in women after menopause,resulting in decreased bone mass and damage to the microstructure of bone tissues. The main clinical manifestations are low back pain,osteoporotic fractures,spinal deformities,and multiple organ dysfunction. PMOP directly leads to high morbidity, high mortality, and a decline in the quality of life. In addition to miss diagnosis, it is often not treated in time. In recent years, significant progress has been made in the research on factors related to the pathogenesis of PMOP. Based on the previous findings in recent years,this article described three major pathogenesis of PMOP, including intestinal flora imbalance,oxidative stress,and abnormal differentiation of bone marrow mesenchymal stem cells (BMMSCs), and analyzed the current status of PMOP treatment, such as syndrome differentiation and treatment,acupuncture and moxibustion,exercise therapy, and external treatment in traditional Chinese medicine (TCM), and basic measures,drug intervention,and physical therapy in western medicine. Among them,drug intervention in western medicine treatment is generally divided into bone resorption inhibitors,bone formation promoters,and other mechanism drugs according to the mechanism of action. This article summarized the specific methods and effects or mechanisms of TCM and western medicine in the clinical treatment of PMOP,which is expected to provide a reference for formulating reasonable health management models and drug treatments in the future.
Keywords:postmenopausal osteoporosis;traditional Chinese medicine treatment;syndrome differentiation and treatment;western medicine treatment;research progress