摘要
目的探讨复方蛇龙胶囊对膜性肾病大鼠肾保护作用及其减轻肾组织细胞凋亡的相关机制。方法60只6周龄SPF级雄性SD大鼠,随机选取10只作为正常组,其余按照改良Border法制备膜性肾病大鼠模型,随机分为模型组、雷公藤多苷片组(7.5mg/kg)和复方蛇龙胶囊低、中、高剂量组(0.375 g/kg,0.75 g/kg,1.5 g/kg),正常组和模型组灌胃给予等体积的生理盐水,每日1次,连续8周。采用全自动生化分析仪检测血清肌酐、甘油三脂、总胆固醇、白蛋白、总蛋白含量;酶联免疫吸附法检测尿微量白蛋白含量;观察肾组织病理学变化;流式细胞术检测肾组织细胞凋亡;实时荧光定量PCR和蛋白免疫印迹法(Western blot,WB)检测肾脏组织Nephrin、Podocin、Bcl-2、Bad的mRNA和蛋白表达水平,WB检测磷酸化蛋白激酶B(p-protein kinase B,p-Akt)蛋白表达水平。结果(1)与正常组比较,模型组血清白蛋白和总蛋白明显降低,总胆固醇、甘油三酯、肌酐和尿微量白蛋白不同程度升高(P<0.05);肾小球结构紊乱,炎性细胞浸润,纤维组织沉积,肾小球基底膜增厚,足细胞损伤;肾组织细胞凋亡率明显升高(P<0.05);足细胞标志分子Nephrin和Podocin表达明显下降(P<0.05);p-Akt蛋白与Bcl-2 mRNA和蛋白表达水平明显降低,Bad的mRNA和蛋白表达水平明显升高(P<0.05);(2)与模型组比较,各给药组白蛋白和总蛋白明显升高,总胆固醇、甘油三酯、肌酐和尿微量白蛋白明显降低(P<0.05);肾组织形态明显改善;细胞凋亡率明显降低(P<0.05);Nephrin和Podocin mRNA和蛋白表达水平明显升高(P<0.05);p-Akt蛋白与Bcl-2 mRNA和蛋白表达明显升高,Bad mRNA和蛋白表达明显降低(P<0.05)。结论复方蛇龙胶囊对膜性肾病大鼠具有明显的肾损伤保护作用,其机制可能与磷脂酰肌醇-3-激酶/蛋白激酶B信号通路抑制肾组织足细胞凋亡有关。
Objective To study the protective effect and possible mechanism of Compound Shelong Capsule on kidneys of membranous nephropathy rats model and renal podocyte apoptosis. Methods Sixty 6-week-old male Sprague-Dawley rats were randomly divided into normal group and model group. Rats in the cationic bovine serum albumin-induced model group were randomly divided into model group, tripterygium glycoside tablets group, and low-dose group, medium-dose group, and high-dose group(0.375 g/kg, 0.75 g/kg, 1.5 g/kg), rats in the normal group and the model group with an equal volume of distilled water, once a day, for eight consecutive weeks. After drug administration, serum creatinine(SCr), triglycerides(TG), total cholesterol(TC), albumin(ALB), total protein(TP) and urinary microalbumin(U-mALB) were detected, the methods of hematoxylin and eosin(HE) staining, Masson’s trichrome staining and transmission electron microscopy were used for observing the pathological changes. Flow cytometry was used to detected cell apoptosis. Real time fluorescent quantitative polymerase chain reaction(Real-time PCR) and western blot were used to detect Nephrin, Podocin, Bcl-2, Bad mRNA and protein expression levels. Western blot was used to detect p-Akt level. Results(1)Compared with the normal control group, the levels of ALB and TP in the model group were significantly decreased, the levels of TC, TG, SCr and U-mALB were increased(P<0.05), disordered glomerular structure, infiltrated inflammatory cells, excessively deposited collagen fibers, thickened basement membrane, extensively fused podocyte foot processes and podocyte injury. Renal tissue cell apoptosis rate was significantly upregulated(P<0.05), podocyte markers Nephrin and Podocin expression levels were significantly downregulated(P<0.05), p-Akt, Bad expression significantly increased, Bcl-2 expression significantly decreased(P<0.05).(2)Compared with the model group, the levels of TC, TG, SCr and U-mALB were significantly decreased in each treated groups(P<0.05), while the levels o
作者
贾世艳
仲启明
司瑞花
范晓阳
严文允
刘光珍
JIA Shiyan;ZHONG Qiming;SI Ruihua;FAN Xiaoyang;YAN Wenyun;LIU Guangzhen(College of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,China)
出处
《环球中医药》
CAS
2022年第10期1788-1795,共8页
Global Traditional Chinese Medicine
基金
山西省重点研发计划重点项目(201603D3113023,201703D421024)
山西省科技攻关项目(2016ZD0307)
山西省晋药综合开发利用协同创新中心项目(2017-JYXT-32)。
