摘要
目的:通过研究加味升降散对膜性肾病(MN)模型大鼠Notch通路信号分子及下游凋亡相关基因的影响,探讨其抑制足细胞凋亡的分子机制。方法:将大鼠随机分为正常组,模型组,加味升降散组和贝那普利组,每组10只。采用尾静脉注射阳离子化牛血清白蛋白(C-BSA)建立MN大鼠模型,加味升降散组和贝那普利组分别以27.3,0.001 g·kg^(-1)灌胃(ig),正常组、模型组ig等体积生理盐水,连续干预4周。采用双缩脲法检测大鼠24小时尿蛋白(UTP)水平,实时荧光定量聚合酶链式反应(Real-time PCR)和免疫组化(IHC)检测大鼠肾组织中足萼蛋白(Podocalyxin),足细胞裂孔膜蛋白(Nephrin),膜蛋白(Podocin),突触足蛋白(Synaptopodin)mRNA及蛋白表水平,原位末端标记法(TUNEL)检测大鼠肾组织细胞凋亡率,蛋白免疫印迹法(Western blot)检测大鼠肾组织中Notch1,Hes1,B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax),Bcl-2蛋白表达水平。结果:与正常组比较,模型组大鼠UTP水平显著升高,肾组织细胞凋亡明显增多,足细胞标志蛋白Podocalyxin,Nephrin,Podocin,Synaptopodin mRNA和蛋白表达水平明显下降,Notch1,Hes1,Bax蛋白表达量明显增多,Bcl-2蛋白表达量明显减少(P<0.05);与模型组比较,加味升降散和贝那普利组大鼠UTP水平明显下降,肾组织细胞凋亡明显减少,足细胞标志蛋白Podocalyxin,Nephrin,Podocin,Synaptopodin mRNA和蛋白表达水平明显升高,Notch1,Hes1,Bax蛋白表达量明显减少,Bcl-2蛋白表达量明显升高(P<0.05)。结论:加味升降散可通过抑制Notch信号通路,减少大鼠肾组织足细胞凋亡,减轻MN大鼠肾损伤。
Objective:The purpose of this article was to observe the effect of modified Shengjiangsan on podocyte apoptosis in membranous nephropathy(MN)rats,to explore the molecular mechanism of its treatment of MN and to provide experimental basis for its clinical application. Method: The MN rat model was established by injection of cationic bovine serum albumin into the tail vein of rats. The successfully modeled rats were then randomly divided into model group(equal volume of normal saline),modified Shengjiangsan group(27.3 g·kg^(-1))and benazepril group(10 mg·kg^(-1)),with corresponding drug dosage once a day for 4 weeks of continuous intervention. After drug administration,the 24-hour urine protein(UTP)was detected. Real time fluorescent quantitative polymerase chain reaction(Real-time PCR)and immunohistochemical(IHC)methods were used to detect Podocalyxin,Nephrin,Podocin,Synaptopodin mRNA and protein expression levels in rat kidney tissue. terninal-deoxynucleoitidyl transferase medsated nick and labeling(TUNEL)method was used to detect cell apoptosis rate in rat kidney tissue, and Western blot was used to detect Notch1, Hes1, B lymphoblastoma-2(Bcl-2)associated X protein(Bax),and Bcl-2 protein expression levels in rat kidney tissue.Result: Compared with the normal group,UTP in the model group increased significantly,renal tissue cell apoptosis increased significantly,podocyte marker proteins podocalyxin,Nephrin,Podocin,Synaptopodin mRNA and protein expression levels decreased significantly, and Notch1, Hes1, Bax protein expression increased significantly,and Bcl-2 protein expression was significantly reduced(P<0.05). Compared with the model group,UTP levels in MN rats were significantly reduced in modified Shengjiangsan and benazepril groups,with reduced rate of renal cell apoptosis,increased mRNA and protein expression levels of podocalyxin,Nephrin,Podocin,and Synaptopodin in renal tissue,decreased Notch1,Hes1,Bax protein expression,and increased Bcl-2 protein expression(P<0.05). Conclusion: Modified Shengjiangsan
作者
高飞
王泽泽
杨冰
杨凤文
檀金川
GAO Fei;WANG Ze-ze;YANG Bing;YANG Feng-wen;TAN Jin-chuan(Hebei University of Chinese Medicine,Shiiazhuang 050200,China;Hebei Provincial Hospital of Chinese Medicine,Shijiazhuang 050011,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第4期30-36,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
河北省自然科学基金项目(2019423037)
河北省中医药管理局项目(2018024)。
