慢肾康宁方对慢性肾衰大鼠肾组织氧化应激状态及Caspase-3,JNK,p66Shc蛋白表达影响  

Effects of Manshenkangning Fang on oxidative stress state and expression levels of Caspase-3,JNK and p66Shc in rat kidneys of chronic renal failure

作　　者：何铃[1] 朱诗平[2] 金鑫[1] 聂玲辉[1] 

机构地区：[1]暨南大学医学院中医系,广东广州510632 [2]暨南大学附属第一医院中医科,广东广州510630

出　　处：《暨南大学学报（自然科学与医学版）》2012年第6期559-564,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30287165);广东省自然科学项目(7005956);广东省建设中医药强省科研课题(2009173)

摘　　要：目的:观察中药慢肾康宁方对腺嘌呤诱发的慢性肾衰大鼠肾脏氧化应激状态及p66Shc,Caspase-3和JNK蛋白的影响。方法:60只wistar大鼠随机分成正常组,模型组,氯沙坦组,中药慢肾康宁方低、中、高质量分数组,除正常组给予生理盐水灌胃外,其余各组使用腺嘌呤灌胃复制慢性肾衰大鼠模型,时间是21d。检测模型建立成功后,正常组和模型组给予生理盐水灌胃,其余各组对应给予等量药物进行治疗,疗程为28d。实验结束后,检测各组大鼠血清尿肌酐(Scr)和尿素氮(BUN)的含量,采用化学比色法测定肾组织中超氧化物歧化酶(SOD)和丙二醛(MDA)的含量,提取肾组织蛋白,通过Western blotting进行p66Shc,Caspase-3和JNK蛋白的检测。结果:造模成功后,模型组Scr及BUN水平明显升高,显著高于正常组(P<0.01),SOD水平降低,MDA活性升高,与正常组相比差异具有统计学意义(P<0.05),肾脏组织内Caspase-3,JNK,p66Shc及其磷酸化位点p66Shc-ser36表达量明显上升(P<0.05)。经药物治疗后,中药低中高剂量组及氯沙坦组Scr和BUN均明显降低(P<0.01),SOD活性升高,中药高剂量组和氯沙坦组升高明显(P<0.05),其余治疗组具有升高趋势,MDA含量明显降低,氯沙坦组和中药高剂量降低趋势明显(P<0.05),其余各组存在降低趋势,Caspase-3,JNK,p66Shc及磷酸化蛋白p66Shc-ser36表达量较模型组相比均呈现出下降趋势(P<0.05)。结论:中药慢肾康宁方高剂量治疗组能够降低慢性肾衰大鼠血清Scr,BUN水平,改善肾间质纤维化程度,作用的机制可能与提高SOD水平,降低MDA表达,改善肾脏氧化应激状态及降低Caspase-3,p66Shc和JNK蛋白表达水平相关。Aim：To observe the effects of Manshenkangning Fang on the oxidative stress state and expression levels of Caspase-3,JNK and p66Shc proteins in chronic renal failure rats kidney. Methods： 60 wistar rats were randomly divided into 6 groups： normal group,model group,losartan group,traditional Chinese medicine low,medium and high dose groups.The normal group was administered intragastrically with 250 mg/kg·d saline while other groups with the same amount adenmine for 21 days.After the model was successfully established,the normal and model groups were administrated intragastrically with saline and other treatment groups with drugs for another 28 days.Then the serum urine cratinine（Scr） and blood urea nitrogen（BUN） was detected and superoxide dismutase（SOD） and malondialdehyde（MDA） content was also detected by automatic biochemistry instrumentation and chemical colorimetric determination.Tissue protein levels of caspse 3,JNK and p66Shc was detected by western blotting analysis. Results： The Scr and BUN levels in model group were significantly higher than normal group（P〈0.01）,and the SOD levels decreased while MDA increased（P〈0.05）.The expression levels of Caspase-3,JNK,p66Shc and phosphorylation sites p66Shc-ser36 were increased significantly（P〈0.05）.The Scr and BUN levels were both decreased after treatment by losartan and traditional Chinese medicine groups（P〈0.01）.The activity of SOD increased in each group while it was significantly in high dose group of traditional Chinese medicine and losartan group（P〈0.05）.But MDA levels decreased in each group and in losartan group and high dose Chinese medicine group it decreased significantly（P〈0.05）.The protein expression levels of Caspase-3,JNK,p66Shc and the phosphorylated protein p66Shc-ser36 were decreased after treatment when compared with model group（P〈0.05）. Conclusion：Traditional Chinese medicine Manshenkangning Fang could reduce chronic renal failure rats serum Scr and BUN levels and improve renal

关 键 词：慢肾康宁方 慢性肾衰 氧化应激 CASPASE-3 P66SHC JNK 

分 类 号：R285.5[医药卫生—中药学]