Abstract：ObjectiveTo explore the efficacy and mechanism of Danshen Zexie decoction in treating the rats with metabolic-associated fatty liver disease （MAFLD）.MethodSixty male SD rats were randomized into control group， model group， essentiale （polyene phosphatidylcholine capsules， 0.144 g·kg-1） group， and low-， medium-， and high-dose Danshen Zexie decoction （1.16， 2.32， and 4.64 g·kg-1， respectively） groups. The rat model of MAFLD was established with high fat diet， and 8-week therapy started at the induction of MAFLD. The serum levels of total cholesterol （TC）， triglyceride （TG）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST）， as well as the liver levels of TC， TG， free fatty acid （FFA）， superoxide dismutase （SOD）， glutathione （GSH）， and malondialdehyde （MDA）， were measured. The pathological changes of liver were observed by hematoxylin-eosin （HE） staining and oil red O staining， and the steatosis score and oil red O staining area were calculated. The level of reactive oxygen species （ROS） in the liver was detected by immunofluorescence. The protein levels of adenosine monophosphate-activated protein kinase （AMPK）， p-AMPK， nuclear factor E2-related factor 2 （Nrf2）， glutamate-cysteine ligase catalytic subunit （GCLC）， glutamate-cysteine ligase modifier subunit （GCLM）， and nicotinamide adenine dinucleotide phosphate： quinine oxidoreductase （NQO1） in the liver were determined by Western blot.ResultCompared with control group， the modeling of MAFLD elevated the levels of TC， TG， ALT， and AST in the serum and TC， TG， FFA， MDA， and ROS in the liver （P<0.01）， lowered the levels of SOD and GSH and down-regulated the protein levels of p-AMPK， Nrf2， GCLC， GCLM， and NQO1 in the liver （P<0.05， P<0.01）. Further， a large number of lipid droplet vacuoles and balloon-like lesions appeared in the hepatocytes， and the steatosis score and oil red O staining area increased in the model group （P<0.01）. Compared with model group， Danshen Zexie decoction lowered the levels of TC， TG， ALT， and AST in the serum and TC， TG， FFA， MDA， and ROS in the liver （P<0.01）， increased the levels of SOD and GSH and up-regulated the protein levels of p-AMPK， Nrf2， GCLC， GCLM， and NQO1 in the liver （P<0.05， P<0.01）. Moreover， the decoction alleviated the degree of liver steatosis （P<0.05， P<0.01）.ConclusionDanshen Zexie decoction protects against MAFLD by activating AMPK/Nrf2 signaling pathway and suppressing oxidative stress.
Abstract：ObjectiveTo explore the possible mechanism of Yiqi Yangyin Huoxue prescription in the prevention and treatment of kidney injury of diabetic kidney disease（DKD）rats based on NOD-like receptor protein 3（NLRP3）/cysteine protease-1（Caspase-1）/gasdermin D （GSDMD）pyroptosis pathway.MethodFifty male SD rats were randomly divided into normal group （8） and modeling group （42）. The modeling group was given a one-time intraperitoneal injection of streptozotocin （STZ） after high-sugar and high-fat diet for 6 weeks to induce the establishment of a DKD rat model. After successful modeling， the rats were randomly divided into model group， valsartan group （8.33 mg·kg-1）， and Yiqi Yangyin Huoxue prescription low-dose and high-dose group （11，22 g·kg-1）. After continuous gavage for 6 weeks， the fasting blood glucose （FBG）， total cholesterol （CHO）， triglyceride （TG）， blood urea nitrogen （BUN）， serum creatinine （SCr） and 24-hour urine protein quantification （24 h-UTP） were detected in each group of rats. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of kidney tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum interleukin-1β （IL-1β） and interleukin-18 （IL-18） levels. The protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue of rats in each group were determined by Western blot and real-time quantitative polymerase chain reaction （Real-time PCR）.ResultCompared with the conditions in normal group， the levels of FBG， CHO， TG， BUN， SCr， 24 h-UTP and serum IL-1β and IL-18 as well as the protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue in model group were increased （P<0.01）， and the kidney tissue lesions were severe. Compared with the conditions in model group， the levels of FBG， CHO， TG， BUN， SCr， 24 h-UTP and serum IL-1β and IL-18 as well as the protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue in each intervention group were decreased （P<0.05， P<0.01）， and the degree of kidney tissue lesions was improved， with Yiqi Yangyin Huoxue prescription high-dose group showing the optimal effect.ConclusionYiqi Yangyin Huoxue prescription could inhibit pyroptosis by regulating the NLRP3/Caspase-1/GSDMD pathway， and thus relieve the inflammatory response of DKD rats and alleviate the pathological damage of the kidneys.
Abstract：ObjectiveTo observe the effects of Hoveniae Semen， Flos Puerariae and their combinations on acute alcoholic liver disease and provide a scientific basis for the drug use in clinical practice and the research on other alcoholic diseases.MethodThe acute alcoholic liver injury model of mice was established by one-time gavage with 56% （V/V） Hongxing Erguotou liquor （12 mL·kg-1）. One hundred and twenty male ICR mice were randomly assigned into blank group， model group， silybin group， Flos Puerariae group， Hoveniae Semen group， and Flos Puerariae-Hoveniae Semen combination groups （ratios of 1∶1， 1∶2 and 2∶1， respectively）， with 15 mice in each group. Each group was administrated with 10 mL·kg-1 corresponding preventive drugs for 3 days by gavage. Except the blank group， the other groups were given Erguotou liquor by gavage at 12 mL·kg-1. The mice were sacrificed 12 h after drinking for the observation of liver function and oxidative stress. The pathological changes of liver were observed via hematoxylin-eosin （HE） staining. Western blot was employed to determine the expression levels of proteins in the Kelch-like Ech-associated protein 1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA levels of related genes.ResultCompared with control group， the modeling elevated the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and alkaline phosphatase （ALP） in the serum and the content of malondialdehyde （MDA） and reactive oxygen species （ROS） in liver tissue （P<0.01） and decreased the activities of glutathione （GSH） and superoxide dismutase （SOD） （P<0.01）. The mRNA and protein expressions of Keap1 were significantly increased （P<0.05，P<0.01）， mRNA and protein expressions of Nrf2 were significantly decreased （P<0.01）. Compared with model group， Flos Puerariae-Hoveniae Semen 2∶1 lowered the levels of ALT， AST and ALP in the serum （P<0.01） and MDA and ROS in the liver （P<0.01） and increased the activities of GSH and SOD （P<0.01）. Moreover， it alleviated the hepatic steatosis injury， up-regulated mRNA and protein levels of Nrf2 （P<0.01） and down-regulated the mRNA and protein levels of Keap1 （P<0.01）.ConclusionFlos Puerariae， Hoveniae Semen and their combinations may exert the pre-protective effect on acute alcoholic liver injury in mice by regulating the Keap1/Nrf2/ARE pathway in the liver and restoring the liver oxidative balance destroyed by ethanol to inhibit the development of alcoholic liver disease .
Keywords：Flos Puerariae;Hoveniae Semen;acute alcoholic liver injury;antioxidation;Kelch-like Ech-associated protein 1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway
LI Weijie,MAO Xia,LIU Yudong,WANG Kexin,ZHANG Yanqiong,LIN Na
Abstract：ObjectiveTo systematically explore the roles and contributions of the sovereign drug Gypsum Fibrosum contained in Baihu Guizhitang （BHGZT） against rheumatoid arthritis （RA） with heat syndrome from property-efficacy association by an approach integrating transcriptomics with gene regulatory network analysis.MethodA total of 20 male Lewis rats were randomly assigned into 4 groups： a control group （n=5）， a group of adjuvant-induced arthritis rat model with heat syndrome （AIA-H， n=5）， an AIA-H + BHGZT group （BHGZT， 21.4 g·kg-1， n=5）， and an AIA-H + BHGZT without Gypsum Fibrosum group （BHGZT-GYP， 10.7 g·kg-1， n=5）. We combined the gene expression profiling based on AIA-H rat model and "disease-gene-drug effective target" network analysis to predict the major function of Gypsum Fibrosum contained in BHGZT against RA with heat syndrome. Furthermore， in vivo experiments with the AIA-H rat model were performed to validate the therapeutic effects on the severity of arthritis based on the representative images of arthritis， limb diameter， infrared thermography， pain thresholds， and joint injury， as well as at the level of immunity-inflammation imbalance. Oil Red O staining was employed for the differentiation of 3T3-L1 pre-adipocytes in the AIA-H rats treated by BHGZT， BHGZT-GYP， and GYP.ResultGene expression profiling and network analysis demonstrated that Gypsum Fibrosum mainly regulated the energy metabolism disorders and the immunity–inflammation imbalance during the development and progression of RA. In vivo experiments showed that both BHGZT and BHGZT-GYP reduced the disease severity of AIA-H rats （P<0.01） by relieving joint redness and distortion， decreasing arthritis score and limb diameter， elevating pain thresholds， alleviating joint erosion， joint inflammation， and bone destruction （P<0.05）. Notably， BHGZT outperformed BHGZT-GYP （P<0.05）. Both BHGZT and BHGZT-GYP inhibited the pathological changes and decreased the indexes of thymus and spleen （P<0.05）， and down-regulated the expression of inflammatory cytokines including Toll-like receptor 4 （TLR4）， tumor necrosis factor-alpha （TNF-α）， interleukin-6 （IL-6）， IL-12， IL-1β， and IL-18 （P<0.05）. In addition， BHGZT and GYP significantly inhibited the adipogenic differentiation of 3T3-L1 cells， with the performance superior to that of BHGZT-GYP.ConclusionThe sovereign drug Gypsum Fibrosum contained in BHGZT played a crucial role in reversing energy metabolism disorders and immunity-inflammation imbalance， which may be associated with its cold property and function of clearing heat and purging fire.
Keywords：Baihu Guizhitang;Gypsum Fibrosum;heat arthralgia;association of property and efficacy;energy metabolism;immunity-inflammation balance
CHEN Zhengtao,LIANG Qingzhi,ZHANG Yuan,YANG Yan,WANG Mengping,XIE Chunguang,GAO Hong
Abstract：Diabetic nephropathy （DN） is one of the serious and common microvascular complications of diabetes mellitus （DM） and the main cause of end-stage renal disease （ESRD）. Endoplasmic reticulum stress （ERS） is a common stress defense mechanism in eukaryotic cells. In the ERS state， cells activate the unfolded protein response （UPR） to enhance the folding of unfolded proteins and the degradation of misfolded proteins， so as to restore the normal physiological function of the endoplasmic reticulum and avoid cell damage. However， excessive or chronic persistent ERS can induce apoptosis， inflammation， oxidative stress and other pathways to eventually cause cell damage. In recent years， a large number of studies have confirmed that ERS is closely associated with the occurrence and development of DN. In the case of DN， ERS is involved in the damage or protection of podocytes， glomerular mesangial cells， renal tubular epithelial cells， and glomerular endothelial cells. The regulation of ERS has become one of the hotspots in the prevention and treatment of DN and has received extensive attention in the field of traditional Chinese medicine. This paper systematically expounds the role of ERS in the occurrence and development of DN and summarizes the ERS-targeted regulation of DN by traditional Chinese medicine， with a view to providing certain research ideas for the prevention and control of DN with traditional Chinese medicine.
Keywords：diabetic nephropathy;endoplasmic reticulum stress;traditional Chinese medicine;renal intrinsic cells
Abstract：Epilepsy is a common nervous system disease， which is characterized by repeated attacks， prolonged and difficult to heal， causing great harm to the patient's body and mind.Antiepileptic drugs bring good therapeutic effect， but also accompanied by a lot of physical and mental damage of toxic side effects.Traditional Chinese medicine has a long history in the treatment of epilepsy. At present， in addition to enriching the cognitive theory of traditional Chinese medicine in the treatment of epilepsy， the research on the cell signal transduction mechanism of traditional Chinese medicine in the treatment of epilepsy from the perspective of molecular biology is developing rapidly.Through literature search at home and abroad， it is found that epilepsy is closely related to cell proliferation， apoptosis， autophagy， inflammatory response， immune response and other pathophysiological processes.At the same time， the modern medical research of traditional Chinese medicine and western medicine shows that there is a direct or indirect relationship between the study on the mechanism of the treatment of epilepsy by monomer Chinese medicine， single Chinese medicine or even compound Chinese medicine.They can inhibit the occurrence of epilepsy， control epilepsy and protect the brain injury caused by epilepsy by regulating the expression of key molecules in the corresponding signaling pathway.In this paper， the research status at home and abroad is summarized as follows：①Tangeretin， ginkgolide B and other drugs can inhibit apoptosis and oxidative stress by activating PI3K/ Akt signaling pathway.②Baicalin，osthol and other drugs can inhibit autophagy by inhibiting mTOR signaling pathway.③Ganoderma polysaccharides， astragaloside and other drugs can reduce cell apoptosis by inhibiting MAPK signaling pathway.④Salidroside，resveratrol and other drugs can inhibit oxidative stress and apoptosis by activating Nrf2/ARE/HO-1 signaling pathway.⑤Curcumin， baicalin and other drugs can reduce inflammation and apoptosis by inhibiting NF-κB signaling pathway.The above summary aims to provide a reference for the in-depth study of the treatment of epilepsy with traditional Chinese medicine， and also provide a new idea for the clinical treatment of epilepsy with traditional Chinese medicine.
Keywords：epilepsy;traditional Chinese medicine;signaingl pathway;research progress