肾衰合剂通过PKA信号通路抑制肾纤维化的实验观察  

Experimental Observation on Shen Shuai Mixture Inhibiting Renal Fibrosis by PKA Signal Channel

作　　者：邹荣[1] 肖方喜[1] 熊飞[1] 陈芳[1] 胡韬韬[1] 黄晓丽[1] 陈菁[1] 梁鸿卿[1] 

机构地区：[1]武汉市第一医院,湖北武汉430022

出　　处：《西部中医药》2015年第6期4-7,共4页Western Journal of Traditional Chinese Medicine

基　　金：2013年湖北省自然科学基金项目(编号2013CFB371);武汉市科学技术局青年科技晨光计划项目(编号20100231037)

摘　　要：目的:观察肾衰合剂对5/6肾切大鼠模型肾纤维化的影响,探讨其防治肾纤维化的可能机制。方法:将30只SD大鼠分成假手术组、模型对照组、中药干预组,假手术组仅行麻醉暴露双肾,剥除肾包膜,不做手术切除,模型对照组、中药干预组5/6肾脏切除建立慢性肾脏病模型,中药干预组在手术1周后每天给予肾衰合剂3 m L灌胃,手术后12周处死大鼠。检测各组大鼠肾功能指标;观察肾脏组织病理变化;免疫荧光法检测αSMA在肾组织中的表达;Western blot检测collagenⅣ、αSMA、CREB、p CREB在肾组织中的蛋白表达水平;Peptag系统检测PKA活化。结果:与假手术组比较,模型组24小时尿蛋白、血尿素氮和血肌酐明显升高(P<0.05),肾脏病理表现为肾小球硬化和肾间质纤维化;蛋白激酶检测提示模型组肾组织PKA明显活化(P<0.05),肾组织中collagenⅣ、αSMA、p CREB蛋白水平均明显升高(P<0.05);中药干预组大鼠24小时尿蛋白、血尿素氮和血肌酐均明显降低(P<0.05),肾小球硬化和间质纤维化程度明显减轻(P<0.05),少量胶原纤维沉积。肾组织中collagenⅣ、αSMA、p CREB蛋白水平均较模型组降低(P<0.05)。结论:肾衰合剂可能通过抑制PKA信号通路减少肾组织中细胞外基质的积聚而抑制肾纤维化。Objective： To explore the possible mechanism of ShenShuai mixture preventing and treating renal fibrosis by observing the influence of ShenShuai mixture on renal fibrosis of 5/6 subtotal nephrectomized rat model. Methods： Thirty SD rats were allocated to sham operation group, model control group and herbal intervention group, the rats in the sham operation group were exposed the kidney under the anaesthesia, to remove renal capsule, the rats in the model control group and herbal intervention group were performed 5/6 nephrectomy to establish the models with chronic renal disease, herbal intervention group received intragastric administration of ShenShuai mixture, 3mL, in one week after the surgery, all the rats were sacrificed in 12 weeks after the surgery. To detect the indexes of renal functions of the rats in all groups; to observe pathological changes of the kidney; to measure the expressions of ctSMA in the renal tissue by immunofluorescence method; to check protein expression levels of collagen IV, c^SMA, CREB and pCREB in renal tissue by Western blot; to examine PKA activation by Peptag system. Results： Compared with sham operation group, 24h urinary protein, blood urea nitrogen, blood creatinine of the model group raised significantly （P〈0.05）, pathological manifestations of the kidney showed glomerular sclerosis and renal interstitial fibrosis; protein kinase suggested that PKA of renal tissue in the model group was activated apparently （P〈0.05）, protein expression levels of collagen IV, ctSMA and pCREB in renal tissue improved notably （P〈0.05）; 24h urinary protein, blood urea nitrogen, blood creatinine of the rats in herbal intervention group decreased obviously（P〈0.05）, the degrees of glomemlar sclerosis and renal interstitial fibrosis were alleviated obviously （P〈0.05）, collagen fibers were accumulated slightly. The levels of collagen IV, ctSMA and pCREB in renal tissue were lower than the model group （P〈0.05）. Conclusion： ShenShuai mixture could prevent rena

关 键 词：肾衰合剂 5/6肾切 肾纤维化 蛋白激酶A 

分 类 号：R285.5[医药卫生—中药学]