Effect and Mechanism of Puerarin on Renal Cell Apoptosis in Rats with Renal Ischemic-reperfusion Injury

ZHU Min-jie; HAO Hai-ying; CHEN Jie; ZHAO Jing-mei; HAO Xiao-juan

doi:10.13422/j.cnki.syfjx.2016230127

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Effect and Mechanism of Puerarin on Renal Cell Apoptosis in Rats with Renal Ischemic-reperfusion Injury

更新时间：2024-01-04

- Effect and Mechanism of Puerarin on Renal Cell Apoptosis in Rats with Renal Ischemic-reperfusion Injury
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 22, Issue 23, Pages: 127-132(2016)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2016230127
  CLC： R285.5
- Published：2016
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ZHU Min-jie, HAO Hai-ying, CHEN Jie, et al. Effect and Mechanism of Puerarin on Renal Cell Apoptosis in Rats with Renal Ischemic-reperfusion Injury[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(23): 127-132.
DOI：

ZHU Min-jie, HAO Hai-ying, CHEN Jie, et al. Effect and Mechanism of Puerarin on Renal Cell Apoptosis in Rats with Renal Ischemic-reperfusion Injury[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(23): 127-132. DOI： 10.13422/j.cnki.syfjx.2016230127.

摘要

目的：研究葛根素对肾缺血再灌注大鼠肾脏组织细胞凋亡的影响，并探讨其可能的作用机制。方法：选取120只实验用大鼠随机分为假手术组、模型组、银杏叶提取物100 mg·kg-1治疗组、葛根素（100，50，25 mg·kg-1）治疗组；采用夹闭双侧肾蒂血管45 min后恢复血流再灌注的方法制备肾缺血再灌注大鼠模型；再灌注后立即通过ip给药，每天1次，疗程2周。治疗完成后，称量体重和肾脏质量并计算肾脏指数，测定血清中尿素氮（BUN），血肌酐（SCr），尿酸（UA）含量，PAS染色法观察肾脏组织形态学变化；末端标记法（TUNEL）染色法观察细胞凋亡状况并计算凋亡指数（AI）；实时荧光定量聚合酶链式反应（Real-time PCR）测定肾脏组织蛋白激酶B（AKT），B细胞淋巴瘤/白血病-2（Bcl-2），Bcl-2相关X蛋白（Bax）mRNA表达，并计算Bax/Bcl-2；蛋白质免疫印迹（Western blot）法检测肾脏组织中半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）蛋白的表达并进行半定量分析；酶联免疫法测定血浆中血浆C-反应蛋白（CRP），肿瘤坏死因子-α（TNF-α），白细胞介素-1β（IL-1β），白细胞介素-6（IL-6），细胞间黏附分子-1（ICAM-1）水平。结果：与模型组比较，葛根素（100，50 mg·kg-1）治疗组大鼠肾脏指数明显降低（P<0.05，P<0.01），血清中BUN，SCr，UA含量明显降低（P<0.05，P<0.01）；葛根素各治疗组肾脏组织病理性形态学变化呈不同程度改善，肾小球细胞凋亡状况呈不同程度好转，均以100 mg·kg-1治疗组效果最为显著，葛根素100，50 mg·kg-1治疗组肾小球细胞AI显著降低（P<0.05，P<0.01），肾脏组织中Bax mRNA表达明显下调且Bcl-2 mRNA表达明显上调（P<0.05，P<0.01），Bax/Bcl-2明显降低（P<0.01），且100 mg·kg-1治疗组AKT mRNA表达显著上调（P<0.01）；Caspase-3蛋白表达量明显降低（P<0.05，P<0.01）。葛根素（100，50 mg·kg-1）治疗组大鼠血浆中CRP，TNF-α，IL-6水平均明显降低（P<0.05，P<0.01），且100 mg·kg-1治疗组IL-1β，ICAM-1含量水平明显降低（P<0.05，P<0.01）。结论：葛根素具有抑制肾缺血再灌注大鼠肾脏组织细胞凋亡、改善肾功能的作用；其作用机制可能与葛根素能够有效上调AKT基因表达、抑制Caspase-3蛋白表达，下调Bax基因表达、上调Bcl-2基因表达，并降低Bax/Bcl-2，降低炎症因子表达、抑制炎症反应有关。

Abstract

Objective: To investigate the effect and mechanism of puerarin on renal cell apoptosis in rats with renal ischemic-reperfusion injury. Method: One hundred and twenty experimental rats were randomly divided into six groups:sham operation group

model control group

ginkgo biloba extract (EGb

100 mg·kg-1) treated group and puerarin 25

100 mg·kg-1 treated groups; the model was established by gripping bilateral renal pedicle vessels for 45 min

the drugs were given immediately after reperfusion once a day for 2 weeks. Two weeks later

the weights were measured

and the renal index (RI) was calculated; the content of blood urea nitrogen(BUN)

serum creatinine(SCr)

uric acid(UA) in serum were determined; the histopathological changes in renal tissues was observed by PAS staining; the apoptosis of renal cells was observed by TUNEL staining

and the apoptosis index (AI) was calculated; the expression levels of AKT

B cell lymphoma/leukemia-2(Bcl-2)

Bcl-2 associated X protein(Bax) mRNA were detected by Real-time PCR

and the Bax/Bcl-2 was calculated; the expression of Caspase-3 in kidney tissues was detected by Western blot

and a semi-quantitative analysis was performed; the levels of C-reactionprotein(CRP)

tumor necrosis factor-α(TNF-α)

interleukin-1β(IL-1β)

IL-6

intercellular adhesion molecule-1(ICAM-1) in plasma were determined. Result: Compared with model control group

the RI in puerarin (100

50 mg·kg-1) treated groups was significantly decreased(P<0.05

P<0.01); the content of BUN

SCr

UA were significantly decreased(P<0.05

P<0.01); the histopathological changes and the apoptosis in puerarin treated groups were significantly alleviated

especially the puerarin (100 mg·kg-1) treated group

and the AI in puerarin (50

100 mg·kg-1) treated groups was significantly decreased(P<0.05

P<0.01); the expression level of Bax mRNA in puerarin (50

100 mg·kg-1) treated groups was significantly decreased

the Bcl-2 mRNA was significantly increased (P<0.05

P<0.01)

the Bax/Bcl-2 was significantly decreased (P<0.01)

and the expression level of AKT mRNA in puerarin (100 mg·kg-1) treated group was significantly increased (P<0.01); the expression of Caspase-3 in puerarin (50

100 mg·kg-1) treated groups was significantly decreased; the level of CRP

TNF-α

IL-6 in plasma of puerarin (50

100 mg·kg-1) treated groups were significantly decreased (P<0.05

P<0.01)

and the IL-1β

ICAM-1 in puerarin (100 mg·kg-1) treated group was significantly decreased (P<0.05

P<0.01). Conclusion: Puerarin had effects in depressing renal cell apoptosis and improving renal function of rats with renal ischemic-reperfusion injury

which may be related to up-regulation of the expression of AKT and Bcl-2 gene

inhibition of the expression of caspase-3 protein

down-regulation of the expression of Bcl-2 gene and reduction of Bax/Bcl-2 and inflammatory reaction.

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