祛风通络方对阿霉素肾病大鼠肾小球α-actinin-4和Desmin表达的干预作用及机理研究

Effect and Mechanism of Qufeng Tongluo Recipe on Expressions of α-actinin-4 and Desmin in Adriamycin-induced Nephropathy Rats

目的:观察祛风通络方对阿霉素肾病(AN)大鼠肾小球滤过屏障及α-actinin-4和Desmin的影响,探讨"祛风通络方"治疗MCN、改善蛋白尿的作用机理,同时对"风伏肾络"的病机理念进行理论的探讨和阐述,为"祛风通络方"的临床应用提供分子原子依据。方法:86只雄性清洁级Sprague-Dawley(SD)健康大鼠随机分为空白对照组(A组)和模型组(B组)、祛风组(祛风通络方、C组)、激祛组(激素+祛风通络方、D组)和激素组(E组)。分别给予相应干预,再次于不同时间测24h蛋白尿及相关生化指标,同时观察大鼠一般情况,肾小球滤过屏障超微结构变化,同时免疫组化染色结果判定[1],测定各组大鼠肾脏组织中α-actinin-4和Desmin的阳性面积百分比,及肾小球中着色的阳性物质光密度值,以判断表达和分布变化。并分析α-actinin-4和Desmin表达变化与尿蛋白的关系,两者之间的关系等。结果:造模成功时与空白对照组比较,模型组大鼠出现:超微结构显示GBM大致正常,足突间距增宽、融合甚至脱落、消失,α-actinin-4和Desmin在足细胞表达上调,高于空白对照组(Pn均<0.01)。α-actinin-4总体量没有显著变化(Pn>0.05),Desmin总体量明显上调(Pn<0.01)。干预结束时与模型组比较:各干预组超微结构病变均有不同程度改善,足突结构较前清晰,各干预组肾小球足细胞α-actinin-4蛋白和Desmin蛋白平均光密度值表达下调,D组减少最明显(Pn<0.01),C组和E组无显著差异(Pn>0.05)。各干预组分布较前规律,差异有统计学意义(Pn<0.05)。结论:1)祛风通络方可以降低阿霉素肾病大鼠尿蛋白排泄,且与激素联用效果更佳,表明其可以改善大鼠低蛋白状态、脂质代谢。2)祛风通络方可以改善α-actinin-4和Desmin的表达分布,可能是该方实现修复足细胞骨架的结构和功能并抑制足细胞进一步损伤的机制之一。

Objective ：To investigation the expressions of a - actinin - 4 and Desmin in adriamycin - induced ne- phropathy（AN） rats and to further explore the effects of Qufeng Tongluo Recipe on the expressions of a - actinin -4 and Desmin in filtration barrier of kidney glomerulus, expecting to provide evidence for its clinical application. And to discuss the pathogenesis theory about pathogenic wind hiding in kidney collaterals in chronic kidney disease（CKD）. Methods ：86 rats with adriamycin nephrophthy were randomly divided into the normal control group（ A, n = 24） , the nephropathy mod- el group （ B）, the Qufeng group （ C）, Qufeng and prednisone group （ D）, prednisone group （E）. With treatment being given respectively, the renal tissue samples were taken out in each group. The distribution and quantity expressions of a - acti- nin - 4 and Desmin were examined by immunohistochemisty. And the uhrastructure and expressions of a - actinin - 4 andDesmin were examined by electron microscope observation. Results ： After adriamycin injection, the expressions of a - actinin-4 and Desmin in the podocyte were up- regulated（ P 〈0. 01 ）. The quantity of a- actinin-4 did not change （P 〉 0. 05） while Desmin increased（ P 〈 0. 01 ）. At the end of interference, the ultrastructure in C, D and E groups came to improve and the average optical expressions of a - actinin - 4 and Desmin were decreased, especially that in D group （P 〈 0. 01 ） and significant deviation was not observed between C and E groups（ P 〉0. 05）. Distributions of a - actinin- 4 and Desmin in C, D and E groups came regular（ P 〈 0. 05）. Conclusion：The abnormal expression and distri- bution of a- actinin - 4 and Desmin are the molecular mechanism that leads to the occurrence and development of pro- teinuria in AN rats. The improvement of the expression and distribution of a- actinin - 4 and Desmin in GBM in glomer- ulus is likely caused by the effects of Qufeng Tongluo Recipe on nephritic syndrome. It is proved that hepathogenesis the- ory about pathogenic wind hiding in kidney collaterals is accordance with the clinical reality and composing prescriptions and treatment of Qufeng Tongluo is reliable in CKD.