复方积雪草防治局灶节段性肾小球硬化模型大鼠足细胞损伤的实验研究

Experimental Research of the Protection of Centalla Asiatica Compound for Podocyte Injury in Focal Segmental Glomerulosclerosis Rats

目的:探讨复方积雪草对局灶节段性肾小球硬化(focal segmental glomerulo sclerosis,FSGS)模型大鼠足细胞裂孔膜蛋白nephrin、podocin表达的影响,阐述其延缓肾小球硬化的部分分子生物学机制。方法:采用左肾摘除+阿霉素重复静脉注射方法建立FSGS模型。复方组分别予高中低剂量复方积雪草膏剂灌胃;对照组予苯那普利混悬液;于第8周末留取24h尿蛋白定量,检测血肌酐(Scr)、尿素氮(BUN)、胆固醇(TC)、三酰甘油(TG)和血浆蛋白(Alb);留取右肾标本,光镜标本行HE染色;用免疫组化法观察裂孔膜蛋白nephrin和podocin表达;RT-PCR法检测肾组织nephrin和podocinmRNA表达。结果:各复方组24h尿蛋白定量及血Scr、BUN、TC、TG较模型组有显著改善,且与西药组疗效相似。RT-PCR及免疫组化结果显示模型组足细胞裂孔膜蛋白表达较正常组明显降低(P<0.05)。而各治疗组表达较模型组均有明显增加(P<0.05),且与正常组比较差异无统计学意义(P>0.05)。结论:复方积雪草能通过上调模型大鼠肾小球内nephrin和podocin分子表达,减轻足细胞损伤,延缓肾小球硬化。

Objective：To investigate the altered expression level of slit diaphragm （SD） protein in focal segmental glomerulosclerosis （FSGS） rats after being treated by centalla asiatica compound（CAC） and to illuminate the CAC’s effect of delaying glomerulosclerosis in molecular level. Methods：The FSGS rat models were made by unilateral nephrectomy plus adriamycin（ADR） injection twice.The CAC-treated rats were respectively treated with low、medium and high dosage of CAC unguentum while the control group was treated with benazepril suspension. At the end of the model-establishing process, the 8th week, the 24h urinary proteinthe and the serum levels of Scr,Bun,Tch,TG and Alb were detected. The cortex tissue in the right kidney was preserved to observe the pathological changes by HE staining through light microscope and to detect the expression of nephrin and podocin in mRNA and protein levels using respectively RT-PCR and immunohistochemistry methods. Results：Compared with the model rats, the levels of 24 h urinary protein、Scr、Bun、Tch and TG were improved significantly in all CAC-treated groups,and there was no marked difference between the CAC groups and the benazepril group. The result of RT-PCR and Immunohistochemistry showed that the nephrin and podocin expression was obviously slight in model group than that in normal group（P〈0.05）.And the expression in all treatment groups were increased markedly in contrast with the model group（P〈0.05）, but with no significant difference contrasted to the normal group（P〈0.05）. Conclusion：The findings suggested that the CAC can protect the podocyte from being injuried and has the better efficiency of anti-glomerulosclerosis in FSGS model rats which probably is obtained through up-regulating the expression of nephrin and podocin .