摘要
目的探讨益气活血化湿方及其拆方对被动型Heymann肾炎(PHN)模型大鼠足细胞裂孔膜蛋白α-actinin-4和CD2AP的调节作用。方法取雄性Wistar大鼠,制备经典的PHN模型。将造模成功的大鼠随机分为模型组、环孢素组、益气活血化湿方全方组、益气方组、活血方组和化湿方组,另取正常大鼠为对照组。各药物组分别灌胃给予相应药液,对照组和模型组大鼠灌胃给予等量0.9%NaCl溶液,连续6周。分别于第0、2、4、6周对各只大鼠眼眶静脉丛采血,代谢笼收集24 h尿液。全自动生化分析仪检测血清样本白蛋白(Alb)、肌酐(SCr)、尿素氮(BUN)水平以及尿液样本24 h尿蛋白定量(UPRO)。干预结束后,取双侧肾脏。HE染色后光镜下观察肾组织形态学变化,透射电镜下观察肾小球基底膜、足细胞及足突的形态学变化。PCR和Western blot检测肾组织中α-actinin-4和CD2AP的mRNA及蛋白表达水平。结果①与对照组相比,模型组大鼠在各时间点UPRO和SCr水平均明显升高(P<0.05),Alb水平明显下降(P<0.05)。与模型组相比,环孢素组第4、6周UPRO水平明显下降(P<0.05),第2、4、6周Alb水平明显下降(P<0.05),但第2、4、6周SCr和BUN水平明显升高(P<0.05)。与模型组相比,全方组仅第6周UPRO水平明显下降(P<0.05)。与环孢素组相比,全方组第2、4、6周SCr和BUN水平明显降低(P<0.05)。②与对照组相比,模型组光镜下观察可见肾小球明显肿大,肾小球基底膜弥漫性增厚,毛细血管襻受压,部分肾小管上皮细胞肿胀;电镜下显示上皮细胞下大量电子致密物沉积,足突广泛融合或消失。与模型组相比,各药物干预组的肾小球病变程度均有减轻。③与对照组相比,模型组α-actinin-4 mRNA和蛋白表达水平显著升高(P<0.05),CD2AP mRNA和蛋白表达水平显著降低(P<0.05)。与模型组相比,环孢素组α-actinin-4 mRNA和蛋白表达水平显著下调(P<0.05),CD2AP mRNA和蛋白表达水平显著上调(P<0.05
Objective To investigate the regulatory effects of Yiqi Huoxue Huashi formula and its decomposed recipes on the podocyte slit membrane proteinα-actinin-4 and CD2AP in rats with passive Heymann nephritis(PHN).Methods Male Wistar rats were used to establish the classical PHN model.The successful modeling rats were randomly assigned into model group,cyclosporine group,Yiqi Huoxue Huashi formula group,Yiqi recipe group,Huoxue recipe group and Huashi recipe group.The normal rats were used as controls.The drug treatment groups were treated with the corresponding drug by intragastric administration,and the control and model groups were treated with equal volume of 0.9%NaCl solution.The treatment course was 6 weeks.Blood was drawn from the orbital venous plexus of rats at weeks 0,2,4 and 6,and 24-hour urine was collected using metabolic cages.The serum levels of albumin(Alb),creatinine(SCr)and urea nitrogen(BUN),and the 24-hour urinary protein quantification(UPRO)were measured by automatic biochemical analyzer.After intervention,the kidneys were collected.The morphological changes of kidney tissues were observed under the light microscope after HE staining,and the morphological changes of glomerular basement membrane,podocytes and podocytic process were observed under the transmission electron microscope.The mRNA and protein expression levels ofα-actinin-4 and CD2AP in kidney tissues were detected by PCR and Western blot.Results①Compared with the control group,the levels of UPRO and SCR increased significantly(P<0.05)and the level of Alb decreased significantly(P<0.05)in the model group at each time point.Compared with the model group,the level of UPRO decreased significantly at weeks 4 and 6(P<0.05),and the level of Alb decreased significantly at weeks 2,4 and 6(P<0.05),while the levels of SCr and BUN increased significantly at weeks 2,4 and 6(P<0.05)in the cyclosporine group.Compared with the model group,the level of UPRO decreased significantly in the Yiqi Huoxue Huashi formula group only at week 6(P<0.05).Compare
作者
雍军
凌春燕
张先闻
王琳
YONG Jun;LING Chunyan;ZHANG Xianwen;WANG Lin(Department of Nephrology,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)
出处
《上海中医药杂志》
2019年第5期69-75,共7页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(81273730)
上海中医药大学附属龙华医院重点专科建设项目(YW001.003)
上海市卫计委进一步加快中医药事业发展三年行动计划项目[ZY(2018-2020)-ZYBZ-02]
