三焦祛湿方对膜性肾病小鼠足细胞中α-actinin-4、Podocalyxin表达的影响

Effects of Sanjiao Qushi Decoction on the Expression of α-actinin-4 and Podocalyxin in Podocytes of Mice with Membranous Nephropathy

目的:探讨三焦祛湿方对膜性肾病小鼠肾组织足细胞α-actinin-4和Podocalyxin表达的影响及其对肾脏保护的可能作用机制。方法:雌性BALB/c小鼠共50只,随机分成正常组10只与造模组40只。造模组小鼠尾部静脉注射阳离子化牛血清白蛋白(C-BSA)建造实验模型,模型复制成功小鼠随机分成模型组、三焦祛湿方组及盐酸贝那普利组,分别给予各治疗组3.71 g/kg、0.001 3 g/kg药物灌胃,1日1次,持续4周。治疗结束,留取小鼠尿液测定尿蛋白水平(24 h-UTP),腹主动脉采血测定血尿素氮(BUN)、血清肌酐(Scr)、总蛋白(TP)、白蛋白(Alb)、三酰甘油(TG)、总胆固醇(TC)生化指标,肾组织标本经苏木素-伊红(HE),马松(Masson),过碘酸-六胺银(PASM)着色,使用光镜观测各组肾组织病理变化,经免疫荧光测定各组肾标本IgG表达情况。蛋白免疫印迹法(Western blot)观测小鼠肾标本α-actinin-4、Podocalyxin蛋白表达情况。结果:与正常组相比,模型组小鼠24 h-UTP、TC、TG水平明显上升(P<0.05),TP、Alb水平明显下降(P<0.05),各组小鼠BUN、Scr水平相比无明显变化,差异无统计学意义(P>0.05);α-actinin-4和Podocalyxin蛋白表达明显降低(P<0.05);与模型组比较,各给药组小鼠24 h-UTP、TC、TG明显下降(P<0.05),TP、Alb明显升高(P<0.05);α-actinin-4和Podocalyxin蛋白表达明显增加(P<0.05)。结论:三焦祛湿方对膜性肾病小鼠肾脏具有显著的保护作用,其机制可能与增加足细胞骨架蛋白α-actinin-4和电荷蛋白Podocalyxin表达有关。

Objective:To investigate the effect of Sanjiao Qushi Decoction on the expression of α-actinin-4 and Podocalyxin in the kidney tissue of mice with membranous nephropathy and its possible mechanism of renal protection.Methods:A total of 50 female BALB/c mice were randomly divided into 10 normal group and 40 model group.The mice in the modeling group were injected with cationized bovine serum Alb umin(C-BSA) into the tail vein to establish an experimental model.The model was successfully replicated.The mice were randomly divided into the model group,the Sanjiao Qushi Decoction group and the benazepril hydrochloride group,and were given each treatment respectively.Groups 3.71 g/kg and 0.001 3 g/kg were given intragastrically,once a day for 4 weeks.At the end of the treatment,the urine of the mice was collected to measure the urine protein level(24 h-UTP),and the abdominal aorta blood was collected to measure the blood urea nitrogen(BUN),serum creatinine(Scr),total protein(TP),Alb umin(Alb),glycerol Triester(TG),total cholesterol(TC) biochemical indexes,kidney tissue samples were stained with hematoxylin-eosin(HE),Masson(Masson),periodic acid-hexamine silver(PASM),and each group was observed by light microscope.The pathological changes of renal tissue,and the expression of IgG in renal specimens of each group was determined by immunofluorescence.The protein expression of α-actinin-4 and Podocalyxin in mouse kidney samples was observed by Western blot.Results:Compared with the normal group,the levels of 24 h-UTP,TC,and TG in the model group were significantly increased(P<0.05),while the levels of TP and Alb were significantly decreased(P<0.05).There was no significant difference in the levels of BUN and Scr in each group(P>0.05).Compared with the model group,the 24 h-UTP,TC and TG of mice in each administration group decreased significantly(P<0.05),TP and Alb were significantly increased(P<0.05);α-actinin-4 and Podocalyxin protein expressions were significantly increased(P<0.05).Conclusion:Sanjiao Qushi Decoction has a significant protective effect on the kidneys of mice with membranous nephropathy,and the mechanism may be related to the increase in the expression of podocyte cytoskeletal protein α-actinin-4 and charge protein Podocalyxin.