葛根素对肾缺血再灌注大鼠肾脏组织细胞凋亡的影响及其机制探讨  

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

作　　者：朱敏杰[1] 郝海英[1] 陈洁[1] 赵京梅[1] 郝晓娟[1] 

机构地区：[1]邯郸市中心医院,河北邯郸056001

出　　处：《中国实验方剂学杂志》2016年第23期127-132,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省自然科学基金项目(08B033)

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^(-1)β(IL^(-1)β),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^(-1)β,ICAM^(-1)含量水平明显降低(P<0.05Objective： 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 25,50,100 mg·kg-1treated 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-2associated 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（ 100mg·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

关 键 词：葛根素 肾脏缺血再灌注 细胞凋亡 机制 

分 类 号：R285.5[医药卫生—中药学]