补肾活血方保护FSGS小鼠足细胞的实验研究

Experiment research of nourishing kidney and activating blood recipe to protect podocytes in focal segmental glomerulosclerosis mice

[目的]观察补肾活血方对局灶性节段性肾小球硬化(FSGS)小鼠足细胞骨架蛋白desmin,足细胞裂孔膜蛋白nephrin及肾母细胞瘤wt1的影响,探讨此组方保护足细胞的作用机制。[方法]以阿霉素诱导方式完成FSGS肾病模型制备,在1周之后以补肾活血方开展治疗(设为治疗组),时间持续6周。然后再设空白对照组与模型组。针对3组的尿、血及肾组织等进行收集,并检测体质量、血清白蛋白(ALB)、24 h尿白蛋白排泄率(UAER)以及尿白蛋白/尿肌酐比值(UACR)等。通过光镜针对肾组织形态变化情况进行观察分析,以免疫荧光完成小鼠的肾组织nephrin荧光强度测定工作;采用逆转录-聚合酶链反应(RT-PCR)检测3组肾组织当中的desmin、wt1表达水平。[结果]模型组UAER、UACR显著高于空白对照组与治疗组(P<0.01),模型组体质量和ALB显著低于空白对照组和治疗组(P<0.01)。空白对照组nephrin免疫荧光表达具有连续性特征,而模型组则是具有少量点状或者片状特征,较空白对照组和治疗组表达明显减少。模型组当中的wt1 mRNA表达水平明显低于治疗组、空白对照组(P<0.01),而desmin mRNA表达水平明显超过空白对照组、治疗组(P<0.01)。[结论]采用补肾活血组方可以降低FSGS小鼠蛋白尿,同时有可能和wt1、nephrin表达水平上调,以及desmin表达水平下调存在关系。

[Objective] To observe the effect of nourishing kidney and activating blood recipe on focal segmental glomerulosclerosis（FSGS） mice cytoskeleton protein desmin, slit diaphragm protein nephrin and renal tumor wt1, come to discuss the mechanism of the protective effect of podocyte. [Methods] The preparation of the FSGS nephropathy model was completed by the adriamycin induction model, and the treatment（set to the treatment group） was conducted after a week, and the duration of the treatment was six weeks. Then the blank control group and the model group were set up. In view of the three groups of kidney tissues and urine, blood, are collected, and detect the body quality, serum albumin（propagated）, 24 hour（UAER）, and urinary albumin/uric creatinine ratio（UACR）, etc. Through the observation and analysis of the morphological changes of the renal tissue by the lens, the nephrin fluorescence intensity of the mice were determined by immunofluorescence. RT-PCR was used to detect desmin, wt1 mRNA expression levels in three groups of renal tissues.[Results] The UAER and UACR in the model group were significantly higher than that in the control group（P〈0.01）, and the quality of the model group and ALB were significantly lower than the control group and the treatment group（P〈0.01）. In the control group, nephrin immunofluorescence expression was characterized by continuous features, while the model group was characterized by a small number of points or flaky features. The expression of nephrin was significantly reduced in model group than of level of control group and treatment group. The expression levels of wt1 mRNA in the model group were significantly lower than those in the treatment group and the control group（P〈0.01）, and the expression levels of desmin mRNA were significantly higher than that in the control group and the treatment group（P〈0.01）. [Conclusion] The study concluded that the use of nourshing kidney and activating blood recipe group could reduce the renal tissue proteinuria in mice with FSGS, and may be associated with wt1, nephrin expression levels, and desmin expression levels.