Yiqi Huoxue Tongluo Prescription Inhibits Activation of Astrocytes to Relieve Diabetic Neuropathic Pain in Rats via ERK Signaling Pathway

YUAN Huilun; CHENG Kai; CHEN Qian; WANG Lujie; LI Jiajie; WANG Yuqing; JIANG Aijuan

doi:10.13422/j.cnki.syfjx.20231243

您当前的位置：

首页 >

文章列表页 >

Yiqi Huoxue Tongluo Prescription Inhibits Activation of Astrocytes to Relieve Diabetic Neuropathic Pain in Rats via ERK Signaling Pathway

更新时间：2023-11-07

- Yiqi Huoxue Tongluo Prescription Inhibits Activation of Astrocytes to Relieve Diabetic Neuropathic Pain in Rats via ERK Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 23, Pages: 36-46(2023)
- 作者机构：
  
  1.安徽中医药大学中西医结合学院，合肥 230012
  2.安徽省中医药科学院中西医结合研究所，合肥 230012
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20231243
  CLC： R2-0;R33;R587.1;R289
- Published：05 December 2023，
  
  Published Online：11 September 2023，
  
  Received：14 July 2023，
- 稿件说明：
扫描看全文
袁慧伦,程凯,陈前等.基于ERK信号通路抑制星形胶质细胞活化探讨益气活血通络方对糖尿病大鼠神经病理性疼痛的作用[J].中国实验方剂学杂志,2023,29(23):36-46.

YUAN Huilun,CHENG Kai,CHEN Qian,et al.Yiqi Huoxue Tongluo Prescription Inhibits Activation of Astrocytes to Relieve Diabetic Neuropathic Pain in Rats via ERK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):36-46.
袁慧伦,程凯,陈前等.基于ERK信号通路抑制星形胶质细胞活化探讨益气活血通络方对糖尿病大鼠神经病理性疼痛的作用[J].中国实验方剂学杂志,2023,29(23):36-46. DOI： 10.13422/j.cnki.syfjx.20231243.

YUAN Huilun,CHENG Kai,CHEN Qian,et al.Yiqi Huoxue Tongluo Prescription Inhibits Activation of Astrocytes to Relieve Diabetic Neuropathic Pain in Rats via ERK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):36-46. DOI： 10.13422/j.cnki.syfjx.20231243.

摘要

目的

探讨益气活血通络方（YHTP）防治糖尿病神经病理性疼痛（DNP）的作用机制。

方法

体内实验将90只SPF级SD雄性大鼠随机分为空白组、模型组、YHTP低剂量组（2.25 g·kg

-1

）、YHTP中剂量组（4.5 g·kg

-1

）、YHTP高剂量组（9 g·kg

-1

）和甲钴胺组（0.175 mg·kg

-1

）。除空白组外，其余5组采用高脂高糖饮食联合小剂量（35 mg·kg

-1

）腹腔注射链脲佐菌素（STZ）构建糖尿病神经病理性疼痛模型。通过神经电生理检测DNP大鼠坐骨神经传导速度，酶联免疫吸附测定法（ELISA）检测大鼠脊髓组织白细胞介素-6（IL-6）、白细胞介素-1

β（

IL-1

β）

和肿瘤坏死因子-

α（

TNF-

α）

炎症因子水平，实时荧光定量聚合酶链式反应（Real-time PCR）检测大鼠脊髓组织中胶质纤维酸性蛋白（GFAP）和细胞外调节蛋白激酶（ERK）的mRNA表达，蛋白免疫印迹法（Western blot）检测大鼠脊髓组织GFAP和磷酸化细胞外调节蛋白激酶（p-ERK）蛋白表达量，免疫荧光测定大鼠脊髓组织GFAP和p-ERK荧光表达水平。体外实验用100 mmol·L

-1

葡萄糖（GLU）诱导星形胶质细胞（CTX-TNA2）活化，构造高糖环境下神经细胞损伤模型。分为空白组、模型组、含药血清组（10% YQHT）、抑制剂组（10 mol·L

-1

COR）、含药血清＋抑制剂组（10% YHTP+10 mol·L

-1

COR），用ELISA检测CTX-TNA2细胞的促炎因子TNF-

、IL-1

及抗炎因子白细胞介素-10（IL-10）表达水平，Western blot检测CTX-TNA2细胞的GFAP和p-ERK蛋白表达量。

结果

动物实验中，与空白组比较，模型组大鼠机械痛阈值（MWT）、热缩足反应时间（TWL）和神经传导速度显著降低（

0.01），空腹血糖、炎症因子、GFAP和p-ERK蛋白，以及ERK1、ERK2、GFAP mRNA表达量显著升高（

0.01）；与模型组比较，YHTP各剂量组大鼠的MWT、TWL和坐骨神经传导速度显著提高（

0.01），IL-1

、TNF-

、IL-6炎性因子含量显著减少（

0.01），脊髓GFAP、p-ERK蛋白，以及ERK1、ERK2、GFAP mRNA表达水平明显降低（

0.05，

0.01）。体外实验中，与空白组比较，模型组细胞存活率降低，促炎因子及ERK、GFAP蛋白表达量显著增加（

0.01）；与模型组比较，YHTP含药血清组IL-1

、TNF-

炎性因子含量明显降低（

0.05，

0.01），抗炎因子IL-10显著增多（

0.01），ERK、GFAP蛋白表达水平显著减少（

0.01）。

结论

益气活血通络方可能通过抑制ERK信号通路介导的星形胶质细胞活化减轻炎症反应，进而改善DNP。

Abstract

Objective

To investigate the mechanism of Yiqi Huoxue Tongluo prescription （YHTP） in the treatment of diabetic neuropathic pain （DNP）.

Method

Ninety SPF-grade SD male rats were randomized into blank， model， low- （2.25 g·kg

-1

）， medium- （4.5 g·kg

-1

）， and high-dose （9 g·kg

-1

） YHTP， and mecobalamin （0.175 mg·kg

-1

） groups. Except those in the blank group， the rats in the remaining 5 groups were fed with a high-fat and high-glucose diet and subjected to intraperitoneal injection of low-dose （35 mg·kg

-1

） streptozotocin （STZ） to establish the model of DNP. The sciatic nerve conduction velocity in DNP rats was measured by the neurophysiological method， and the levels of interleukin-6 （IL-6）， IL-1

， and tumor necrosis factor-

（TNF-

） were measured by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to measure the mRNA levels of glial fibrillary acidic protein （GFAP） and extracellular signal-regulated kinase （ERK） in the spinal cord. Western blot was employed to measure the protein levels of GFAP and phosphorylated ERK （p-ERK）， and immunofluorescence staining to measure the fluorescence intensity of GFAP and p-ERK in the spinal cord. In the cell experiments， 100 mmol·L

-1

high glucose was used to induce the activation of astrocytes （CTX-TNA2） for the modeling of nerve cell injury. The cells were randomized into the normal， model， drug-containing serum （10% YQHT）， inhibitor ［10 mol·L

-1

corynoxeine （COR）］， drug-containing serum + inhibitor （10% YHTP + 10 mol·L

-1

COR） groups. The levels of pro-inflammatory factors （TNF-

and IL-1

） and the anti-inflammatory factor IL-10 in CTX-TNA2 cells were determined by ELISA， and the protein levels of GFAP and p-ERK in CTX-TNA2 cells by Western blot.

Result

The animal experiments showed that compared with the blank group， the model group presented reduced mechanical withdrawal threshold （MWT）， thermal work limit （TWL）， and nerve conduction velocity， elevated levels of fasting blood glucose， IL-1

， TNF-

， and IL-6， and up-regulated protein levels of GFAP and p-ERK， and mRNA levels of ERK1， ERK2， GFAP （

0.01）. Compared with model group， YHTP increased the MWT， TWL， and sciatic nerve conduction velocity （

0.01）， lowered the levels of IL-1

， TNF-

， and IL-6 （

0.01）， and down-regulated the protein levels of GFAP and p-ERK， and mRNA levels of ERK1， ERK2， GFAP in the spinal cord （

0.05，

0.01）. The cell experiments showed that compared with the blank group， the model group had decreased survival rate， elevated levels of pro-inflammatory factors， and up-regulated protein levels of ERK and GFAP （

0.01）. Compared with the model group， the YHTP-containing serum lowered the levels of IL-1

and TNF-

（

0.05，

0.01）， elevated the level of IL-10 （

0.01）， and down-regulated the protein levels of ERK and GFAP （

0.01）.

Conclusion

YHTP may inhibit the activation of astrocytes by inhibiting the ERK signaling pathway to reduce inflammation and thus relieve DNP.

关键词

糖尿病神经病理性疼痛益气活血通络方细胞外调节蛋白激酶（ERK）信号通路星形胶质细胞

Keywords

diabetic neuropathic painYiqi Huoxue Tongluo prescriptionextracellular signal-regulated kinase （ERK） pathwayastrocytes

references

YANG K，WANG Y，LI Y W，et al.Progress in the treatment of diabetic peripheral neuropathy［J］.Biomed Pharmacother，2022，148：112717.

ELAFROS M A，ANDERSEN H，BENNETT D L，et al.Towards prevention of diabetic peripheral neuropathy：Clinical presentation，pathogenesis，and new treatments［J］.Lancet Neurol，2022，21（10）：922-936.

LIU X，HE J，GAO J，et al.Fluorocitrate and neurotropin confer analgesic effects on neuropathic pain in diabetic rats via inhibition of astrocyte activation in the periaqueductal gray［J］.Neurosci Lett，2022，768：136378.

REN W J，ILLES P.Involvement of P2X7 receptors in chronic pain disorders［J］.Purinergic Signal，2022，18（1）：83-92.

LI D，LIU X，LIU T，et al.Neurochemical regulation of the expression and function of glial fibrillary acidic protein in astrocytes［J］.Glia，2020，68（5）：878-897.

JI R R，DONNELLY C R，NEDERGAARD M.Astrocytes in chronic pain and itch［J］.Nat Rev Neurosci，2019，20（11）：667-685.

MARUTA T，NEMOTO T，HIDAKA K，et al.Upregulation of ERK phosphorylation in rat dorsal root ganglion neurons contributes to oxaliplatin-induced chronic neuropathic pain［J］.PLoS One，2019，14（11）：e0225586.

HUANG S，CHEN Y，JIA Y，et al.Delayed inhibition of ERK and p38 attenuates neuropathic pain without affecting motor function recovery after peripheral nerve injury［J］.Neuropharmacology，2022，202：108835.

DAULHAC L，MALLET C，COURTEIX C，et al.Diabetes-induced mechanical hyperalgesia involves spinal mitogen-activated protein kinase activation in neurons and microglia via N-methyl-D-aspartate-dependent mechanisms［J］.Mol Pharmacol，2006，70（4）：1246-1254.

林逸轩，王帆竞，李金菊，等.益气活血通络方治疗糖尿病周围神经病变疗效观察［J］.中医药临床杂志，2019，31（11）：2135-2138.

马俊龙，唐何勇，王帆竞，等.益气活血通络方对糖尿病神经病理性疼痛大鼠脊髓星形胶质细胞活性及JNK信号通路的影响［J］.中华中医药杂志，2021，36（3）：1397-1401.

江爱娟，申国明，蔡标，等.益气活血通络方通过调控NF-κB信号通路保护db/db小鼠坐骨神经作用机制研究［J］.中国中西医结合杂志，2018，38（8）：985-990.

姚永传，申国明，江爱娟，等.益气活血通络方对糖尿病周围神经病变小鼠JNK信号通路蛋白及炎性因子的影响［J］.北京中医药大学学报，2017，40（11）：923-927.

TANG H Y，WANG F J，MA J L，et al.Acupuncture attenuates the development of diabetic peripheral neuralgia by regulating P2X4 expression and inflammation in rat spinal microglia［J］.J Physiol Sci，2020，70（1）：45.

WANG F，TANG H，MA J，et al.The effect of Yiqi Huoxue Tongluo decoction on spinal cord microglia activation and ASK1-MKK3-p38 signal pathway in rats with diabetic neuropathic pain［J］.Evid Based Complement Alternat Med，2022，2022：2408265.

SUN H，SAEEDI P，KARURANGA S，et al.IDF diabetes atlas：Global，regional and country-level diabetes prevalence estimates for 2021 and projections for 2045［J］.Diabetes Res Clin Pract，2022，183：109119.

ZAKIN E，ABRAMS R，SIMPSON D M.Diabetic neuropathy［J］.Semin Neurol，2019，39（5）：560-569.

SELVARAJAH D，KAR D，KHUNTI K，et al.Diabetic peripheral neuropathy：Advances in diagnosis and strategies for screening and early intervention［J］.Lancet Diabetes Endocrinol，2019，7（12）：938-948.

LI Z，WANG W，MENG F，et al.Analgesic and neuroprotective effects of Baimai Ointment on diabetic peripheral neuropathy［J］.J Ethnopharmacol，2022，292：115122.

SLOAN G，ALAM U，SELVARAJAH D，et al.The treatment of painful diabetic neuropathy［J］.Curr Diabetes Rev，2022，18（5）：e070721194556.

REN Y，XIONG P，ZHONG C，et al.The effect of Gegen Qinlian decoction on clinical prognosis and islet function for type 2 diabetic mellitus：A protocol for systematic review and meta-analysis［J］.Medicine （Baltimore），2021，100（5）：e24210.

CAO S，HOU G，MENG Y，et al.Network pharmacology and molecular docking-based investigation of potential targets of Astragalus membranaceus and Angelica sinensis compound acting on spinal cord injury［J］.Dis Markers， 2022， 2022：2141882.

严威忠，李惠萍，胡剑利.芪归糖痛宁颗粒对糖尿病的临床有效性评价［J］.中药材，2015，38（4）：881-882.

YE G，LIN C，ZHANG Y，et al.Quercetin alleviates neuropathic pain in the rat CCI model by mediating AMPK/MAPK pathway［J］.J Pain Res，2021，14：1289-1301.

LIN X，WANG M，ZHANG J，et al.p38 MAPK：A potential target of chronic pain［J］.Curr Med Chem，2014，21（38）：4405-4418.

KONG F，SUN K，ZHU J，et al.PD-L1 improves motor function and alleviates neuropathic pain in male mice after spinal cord injury by inhibiting MAPK pathway［J］.Front Immunol，2021，12：670646.

LU R，CUI S S，WANG X X，et al.Astrocytic c-Jun N-terminal kinase-histone deacetylase-2 cascade contributes to glutamate transporter-1 decrease and mechanical allodynia following peripheral nerve injury in rats［J］.Brain Res Bull，2021，175：213-223.

LI J，WEI Y，ZHOU J，et al.Activation of locus coeruleus-spinal cord noradrenergic neurons alleviates neuropathic pain in mice via reducing neuroinflammation from astrocytes and microglia in spinal dorsal horn［J］.J Neuroinflammation，2022，19（1）：123.

ARDELEANU V，TOMA A，PAFILI K，et al.Current pharmacological treatment of painful diabetic neuropathy：A narrative review［J］.Medicina （Kaunas），2020，56（1）：25.

CHEN Y L，FENG X L，CHEUNG C W，et al.Mode of action of astrocytes in pain：From the spinal cord to the brain［J］.Prog Neurobiol，2022，219：102365.

DENG S，CHEN X，LEI Q，et al.AQP2 promotes astrocyte activation by modulating the TLR4/NF-κB-p65 pathway following intracerebral hemorrhage［J］.Front Immunol，2022，13：847360.

DONNELLY C R，ANDRIESSEN A S，CHEN G，et al.Central nervous system targets：Glial cell mechanisms in chronic pain［J］.Neurotherapeutics，2020，17（3）：846-860.

LU H J，GAO Y J.Astrocytes in chronic pain：Cellular and molecular mechanisms［J］.Neurosci Bull，2023，39（3）：425-439.

YANG B，LI Y，MA Y，et al.Selenium attenuates ischemia/reperfusion injury‑induced damage to the blood‑brain barrier in hyperglycemia through PI3K/Akt/mTOR pathway‑mediated autophagy inhibition ［J］.Int J Mol Med， 2021， 48（3）：178.

BU X，ZHANG A，CHEN Z，et al.Migration of gastric cancer is suppressed by recombinant newcastle disease virus （rL-RVG） via regulating α7-nicotinic acetylcholine receptors/ERK- EMT［J］.BMC Cancer，2019，19（1）：976.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Mechanism of Dihuang Yinzi in Improving Astrocyte Injury and Glycolysis in AD Mice via PI3K/Akt Pathway

Mechanism of Dihuang Yinzi in Improving Energy Metabolism Disorder and Autophagy Injury of Astrocytes in Brain of AD Mice

Mechanism of Dihuang Yinzi in Improving Astrocyte Injury and Regulating Synaptic Structure and Function in AD Mice

Effect of Zuogui Jiangtang Tongmai Prescription on Astrocyte Injury by AGEs Combined with Oxygen-glucose Deprivation Based on AMPK/mTOR/ULK1 Pathway Related to Autophagy

Related Author

WANG Fengli

SUN Mengjie

YU Hongni

KANG Shenghua

HAN Guanghui

LI Dongyue

ZHEN Weizhe

MA Tao

Related Institution

Dongfang Hospital,Beijing University of Chinese Medicine

School of Traditional Chinese Medicine,Beijing University of Chinese Medicine

Japan Friendship Hospital

Dongfang Hospital，Beijing University of Chinese Medicine

School of Traditional Chinese Medicine， Beijing University of Chinese Medicine

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰