Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy and Dosage of Astragalus in Prescription： Based on SIRT1/p53-mediated Apoptosis

Ying BEN; Tian-ya ZHANG; Jia-xin TIAN; Zhi-hong ZHANG

doi:10.13422/j.cnki.syfjx.20212401

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Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy and Dosage of Astragalus in Prescription： Based on SIRT1/p53-mediated Apoptosis

更新时间：2021-12-14

- Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy and Dosage of Astragalus in Prescription： Based on SIRT1/p53-mediated Apoptosis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 2, Pages: 1-10(2022)
- 作者机构：
  
  河北中医学院中西医结合学院，河北省中西医结合肝肾病证重点实验室，石家庄 050091
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20212401
  CLC： R2-0;R22;R285.5;R289;R33
- Published：20 January 2022，
  
  Published Online：25 October 2021，
  
  Received：03 August 2021，
- 稿件说明：
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贲莹,张天雅,田佳鑫等.基于SIRT1/p53介导的细胞凋亡途径探讨补阳还五汤对糖尿病周围神经病变的治疗作用及方中黄芪用量[J].中国实验方剂学杂志,2022,28(02):1-10.

BEN Ying,ZHANG Tian-ya,TIAN Jia-xin,et al.Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy and Dosage of Astragalus in Prescription： Based on SIRT1/p53-mediated Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):1-10.
贲莹,张天雅,田佳鑫等.基于SIRT1/p53介导的细胞凋亡途径探讨补阳还五汤对糖尿病周围神经病变的治疗作用及方中黄芪用量[J].中国实验方剂学杂志,2022,28(02):1-10. DOI： 10.13422/j.cnki.syfjx.20212401.

BEN Ying,ZHANG Tian-ya,TIAN Jia-xin,et al.Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy and Dosage of Astragalus in Prescription： Based on SIRT1/p53-mediated Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):1-10. DOI： 10.13422/j.cnki.syfjx.20212401.

摘要

目的

通过观察补阳还五汤调节沉默信息调节因子2相关酶1（SIRT1）/p53通路发挥对糖尿病周围神经病变（DPN）大鼠的神经保护作用，探讨其治疗DPN的机制和方中黄芪用量。

方法

90只SD大鼠随机分为正常组，模型组，DPN加含120 g黄芪的补阳还五汤组（BYHW120组），DPN加含60 g黄芪的补阳还五汤组（BYHW60组），DPN加含30 g黄芪的补阳还五汤组（BYHW30组），DPN加

-硫辛酸组（ALA组）。除正常组外，其余各组大鼠均用高糖高脂饲料和腹腔注射链脲佐菌素诱导糖尿病。各用药干预组分别给与相应药物持续干预12周，剂量依次为含120 g黄芪的补阳还五汤15g·kg

-1

·d

-1

，含60 g黄芪的补阳还五汤8.75 g·kg

-1

·d

-1

，含30 g黄芪的补阳还五汤 5.625 g·kg

-1

·d

-1

；

-硫辛酸 60 mg·kg

-1

·d

-1

。正常组给予标准饮食。干预结束后检测机械性痛阈和神经传导速度；取L4-5背根神经节，苏木素-伊红（HE）染色、甲苯胺蓝染色观察病理形态改变，原位末端标记法（TUNEL）染色观察神经细胞凋亡情况；免疫组化和蛋白免疫印迹法（Western blot）检测剪切的半胱氨酸天冬氨酸蛋白水解酶-3（cleaved Caspase-3），SIRT1/p53通路中主要蛋白SIRT1，乙酰化的p53（acetyl-p53），线粒体相关动力蛋白1（Drp1），B细胞淋巴瘤-2（Bcl-2）和Bcl-2相关X蛋白（Bax）的表达。

结果

与正常组比较，模型组血糖显著升高（

0.01），神经传导速度、机械性痛阈均显著降低（

0.01），TUNEL染色阳性细胞百分比，cleaved Caspase-3表达显著升高（

0.01），SIRT1表达显著降低（

0.01），acetyl-p53，Drp1，Bax/Bcl-2均显著升高（

0.01）。与模型组比较，各用药干预组cleaved Caspase-3，acetyl-p53，Drp1，Bax/Bcl-2均显著降低（

0.01）；除BYHW30组外，其余用药干预组神经传导速度、机械性痛阈明显升高（

0.05，

0.01），TUNEL染色阳性细胞百分比明显升高（

0.05，

0.01）；BYHW120组，ALA组SIRT1表达明显升高（

0.05，

0.01）。与BYHW120组比较，BYHW60组，BYHW30组的感觉神经传导速度、机械性痛阈及SIRT1表达明显降低（

0.05，

0.01），cleaved Caspase-3表达显著升高（

0.01）；BYHW30组TUNEL染色阳性细胞百分比，acetyl-p53表达显著升高（

0.01）。

结论

补阳还五汤通过调节SIRT1/p53抑制细胞凋亡，治疗DPN。其治疗作用与方中黄芪的用量有一定关系，120 g黄芪剂量的补阳还五汤抑制p53依赖的细胞凋亡的作用较60 g和30 g黄芪剂量补阳还五汤更明显。

Abstract

Objective

To explore the neuroprotective effect of Buyang Huanwutang （BYHW） on diabetic peripheral neuropathy （DPN） rats by regulating SIRT1/p53 pathway and to clarify the mechanism and the dosage of astragalus in the prescription.

Method

A total of 90 SD rats were randomized into control group， DPN group， DPN + BYHW containing 120 g Astragalus （at 15 g·kg

-1

·d

-1

）（BYHW120 group）， DPN + BYHW containing 60 g Astragalus （at 8.75 g·kg

-1

·d

-1

）（BYHW60 group）， DPN + BYHW containing 30 g Astragalus （at 5.625 g·kg

-1

·d

-1

）（BYHW30 group）， and DPN +

-lipoic acid （at 60 mg·kg

-1

·d

-1

）（ALA group）. Standard diet was given to rats in the control group and high-carbohydrate/high-fat diet and streptozotocin （ip） were used to induce diabetes in rats in other groups. The administration lasted 12 weeks. After the intervention， mechanical pain threshold and nerve conduction velocity were detected. The L4-5 dorsal root ganglions were stained with haematoxylin-eosin （HE） and toluidine blue to observe the pathological changes， and the apoptosis of nerve cells was detected by terminal deoxynucleotidal transferase-mediated dUTP-biotin nick end labeling （TUNEL） assay. Cleaved cysteinyl aspartate-specific proteinase-3 （Caspase-3）， and the main proteins in the SIRT1/p53 pathway， such as silencing information regulator 2 related enzyme 1 （SIRT1）， acetyl-p53， dynamin-related protein 1 （Drp1）， Bcl-2 associated X protein （Bax）， and B-cell lymphoma-2 （Bcl-2）， were detected by immunohistochemistry and Western blot.

Result

Compared with the control group， the DPN group presented increase in blood glucose （

0.01）， decrease in nerve conduction velocity and mechanical pain threshold （

0.01）， rise of the percentage of positive cells （TUNEL assay， the same below） and the expression of cleaved Caspase-3 （

0.01）， drop in the expression of SIRT1 （

0.01）， and elevation of acetyl-p53， Drp1， and Bax/Bcl-2 ratio （

0.01）. Cleaved Caspase-3， acetyl-p53， Drp1， and Bax/Bcl-2 ratio in each administration group decreased as compared with those in the DPN group （

0.01）. Nerve conduction velocity， mechanical pain threshold （

0.05，

0.01）， and the percentage of positive cells （

0.05，

0.01） increased in the administration groups as compared with those in the DPN group except for the BYHW30 group， and BYHW120 group and ALA group showed the increase in SIRT1 （

0.05，

0.01）. Nerve conduction velocity， mechanical pain threshold， and SIRT1 expression were lower （

0.05，

0.01） and expression of cleaved Caspase-3 was higher （

0.01） in the BYHW60 and BYHW30 groups than in the BYHW120 group. The percentage of positive cells and the expression of acetyl-p53 were higher in the BYHW30 group than in the BYHW120 group （

0.01）.

Conclusion

BYHW inhibits apoptosis and exerts therapeutic effect on DPN by regulating the SIRT1/p53 pathway. The therapeutic effect is related to the dosage of Astragalus in the prescription. BYHW containing 120 g Astragalus suppresses p53-dependent apoptosis more significantly than Buyang Huanwutang containing 60 g and 30 g of Astragalus.

关键词

糖尿病周围神经病变补阳还五汤黄芪沉默信息调节因子2相关酶1（SIRT1）/p53通路细胞凋亡

Keywords

diabetic peripheral neuropathyBuyang Huanwutangastragalussilencing information regulator 2 related enzyme 1 （SIRT1）/p53 pathwayapoptosis

references

TESFAYE S.Advances in the management of diabetic peripheral neuropathy［J］.Curr Opin Support Palliat Care，2009，3（2）：136-143.

ANG L，JAISWAL M，MARTIN C，et al.Glucose control and diabetic neuropathy： lessons from recent large clinical trials［J］.Curr Diab Rep，2014，14（9）：528.

YANG C P，LIN C C，LI CI，et al.Cardiovascular risk factors increase the risks of diabetic peripheral neuropathy in patients with type 2 diabetes mellitus：the taiwan diabetes study［J］.Medicine （Baltimore），2015，94（42）：e1783.

DEWANJEE S，DAS S，DAS A K，et al.Molecular mechanism of diabetic neuropathy and its pharmacotherapeutic targets［J］.Eur J Pharmacol，2018，833：472-523.

CHENG C，ZOCHODNE D W.Sensory neurons with activated Caspase-3 survive long-term experimental diabetes［J］.Diabetes，2003，52（9）：2363-2371.

KOBAYASHI M，CHANDRASEKHAR A，CHENG C，et al.Diabetic polyneuropathy，sensory neurons，nuclear structure and spliceosome alterations： a role for CWC22［J］.Dis Model Mech，2017，10（3）：215-224.

ZOCHODNE D W，RAMJI N，TOTH C.Neuronal targeting in diabetes mellitus：a story of sensory neurons and motor neurons［J］.Neuroscientist，2008，14（4）：311-318.

ØSTERGAARD L，FINNERUP N B，TERKELSEN A J，et al.The effects of capillary dysfunction on oxygen and glucose extraction in diabetic neuropathy［J］.Diabetologia，2015，58（4）：666-677.

YOREK M A.Vascular impairment of epineurial arterioles of the sciatic nerve：implications for diabetic peripheral neuropathy［J］.Rev Diabet Stud，2015，12（1）：13-28.

POULOSE N，RAJU R.Sirtuin regulation in aging and injury［J］.Biochim Biophys Acta，2015，1852（11）：2442-2455.

孙洪宽，田园.补阳还五汤加味治疗糖尿病周围神经病变临床观察［J］.北京中医药，2009，28（7）： 541-542.

丁亚琴，吴坚，谢心等.针灸联合补阳还五汤对糖尿病周围神经病变患者神经功能、血清NSE水平的影响［J］.上海针灸杂志，2019，38（8）：865-869.

祖丽华，王心东，秦玖刚.补阳还五汤联合硫辛酸治疗糖尿病周围神经病变临床研究［J］.新中医，2020，52（14）：37-41.

COPPEY L，DAVIDSON E，SHEVALYE H，et al.Progressive loss of corneal nerve fibers and sensitivity in rats modeling obesity and type 2 diabetes is reversible with omega-3 fatty acid intervention： supporting cornea analyses as a marker for peripheral neuropathy and treatment［J］.Diabetes Metab Syndr Obes，2020，13：1367-1384.

BIESSELS G J，BRIL V，CALCUTT N A，et al.Phenotyping animal models of diabetic neuropathy：a consensus statement of the diabetic neuropathy study group of the EASD （neurodiab）［J］.J Peripher Nerv Syst，2014，19（2）：77-87.

马理元，姜劲挺，张伦广，等.逆神经走行方向探寻并摘取大鼠背根神经节的改进方法［J］.中国比较医学杂志，2018，28（3）：95-97.

YAMADA K，YOSHIDA K.Mechanical insights into the regulation of programmed cell death by p53 via mitochondria［J］.Biochim Biophys Acta Mol Cell Res，2019，1866（5）：839-848.

VASEVA A V，MOLL U M.The mitochondrial p53 pathway［J］.Biochim Biophys Acta，2009，1787（5）：414-420.

GREEN D R，KROEMER G.Cytoplasmic functions of the tumour suppressor p53［J］.Nature，2009，458（7242）：1127-1130.

DUBOFF B，FEANY M，GOTZ J.Why size matters-balancing mitochondrial dynamics in Alzheimer's disease［J］.Trends Neurosci，2013，36（6）：325-335.

ITOH K，NAKAMURA K，IIJIMA M，et al.Mitochondrial dynamics in neurodegeneration［J］.Trends Cell Biol，2013，23（2）：64-71.

MCINNES J.Insights on altered mitochondrial function and dynamics in the pathogenesis of neurodegeneration［J］.Transl Neurodegener，2013，2（1）：12.

FRANK S，GAUME B，BERGMANN-LEITNER E S，et al.The role of dynamin-related protein 1，a mediator of mitochondrial fission，in apoptosis［J］.Dev Cell，2001，1（4）：515-525.

BROOKS C，GU W.The impact of acetylation and deacetylation on the p53 pathway［J］.Protein Cell，2011，2（6）：456-462.

ZHOU H，HU S，JIN Q，et al.Mff-dependent mitochondrial fission contributes to the pathogenesis of cardiac microvasculature ischemia/reperfusion injury via induction of mROS-mediated cardiolipin oxidation and HK2/VDAC1 disassociation-involved mPTP opening［J］.J Am Heart Assoc，2017，6（3）：e005328.

ZHOU H，DU W，LI Y，et al.Effects of melatonin on fatty liver disease：the role of NR4A1/DNA-PKcs/p53 pathway，mitochondrial fission，and mitophagy［J］.J Pineal Res，2018，64（1）：e12450-12452.

XIONG H，PANG J，YANG H，et al.Activation of miR-34a/SIRT1/p53 signaling contributes to cochlear hair cell apoptosis：implications for age-related hearing loss［J］.Neurobiol Aging，2015，36（4）：1692-1701.

POULOSE N，RAJU R.Sirtuin regulation in aging and injury［J］.Biochim Biophys Acta，2015，1852（11）：2442-2455.

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