加味当归补血汤对细胞骨架蛋白在阿霉素肾病大鼠表达的影响

Therapeutic Effect of Jiawei Danggui Buxue Tang on Podocyte in Daunomycin-induced Nephropathy

摘要:

目的: 利用阿霉素肾病模型(daunomycin-induce nephropathy, DMN)研究加味当归补血汤通过干预细胞骨架蛋白的表达对肾小球滤过作用所产生的影响.方法: SD大鼠分为正常组、模型组、贝那普利组(1 mg ·kg^-1)和加味当归补血汤组(10 g ·kg^-1).其中模型组、贝纳普利组和加味当归补血汤组使用阿霉素一次性尾静脉注射6 mg ·kg^-1.造模后第2天分别ig给予贝纳普利组和加味当归补血汤组相应的药物,每日1次,共8周.分别在造模第7,28,42,56天留取尿液标本,观察尿白蛋白定量的动态变化;取肾组织进行光镜、免疫组织化学对nephrin,巢蛋白(nestin)和vimentin的表达情况进行分析.结果: 尿白蛋白:在28,42,56 d模型组与正常组比较尿蛋白增加均非常明显,差异有统计学意义(P<0.05);各治疗组与模型组比较明显下降,差异有统计学意义(P<0.05),且加味当归补血汤更显著.肾脏组织:光镜和免疫组织化学结果显示,加味当归补血汤治疗组与贝那普利治疗组和模型组比较系膜细胞增生、肾小管-间质病变情况和肾组织nephrin,nestin和vimentin蛋白的表达情况均减轻,尿白蛋白定量比模型组明显减少,说明肾脏病变程度相比模型组减轻;治疗56 d后肾组织RT-PCR及Wetern blot检测nephrin,nestin和vimentin表达之间存在线性关系;治疗组与模型组比较差异有统计学意义(P<0.05).结论: 加味当归补血汤对于阿霉素肾病模型的作用表现为保护足细胞裂孔膜结构的完整性的同时抑制系膜细胞增生和减轻肾小管-间质损伤,上述作用与改善细胞骨架蛋白的表达呈正相关.

Abstract:

Objective:To study the effect of Jiawei Danggui Buxue Tang(JWDGBXT) on daunomycin-induced nephropathy (DN) by in rats. Method: SD rats were divided into four groups, including normal controls, model group, benazeprilgroup(1 mg · kg^-1) JWDGBXT(10 g · kg^-1). The adriamycin atdose of 6 mg · kg^-1 was used to establish the model. The benepril or JWDGBXT was given by intragastric administration. 7, 28, 42 and 56 days after the treatment, the levels of microdosis proteinuria were tested. The renal tissue was processed for the examinations of light microscopy and immunohistochemistry. Then the renal tissue was assayed by the way of RT-PCR and Wetern blot to observe the level of nephrin, nestin and vimentin. Result: Compared with the normal group, model group had obvilus changes at the 24 h urine protein(P<0.05). Moreover, JWDGBXT could significantly reduce protein uria in rats with daunomycin-induce nephropathy than model group and benepril group(P<0.05). The intercapillary cells had different hyperplasy of the SD rats. The pathological changes of renal tissue at renal tubular and renal interstitial were analyzed. Then, the level of nephrin, nestin and vimentin of the renal tissue was detected by the way of RT-PCR and Western blot. The level of nephrin, nestin and vimentin wa improved. Conclusion: JWDGBXT could decrease the level of proteinuria, protecte the podocytes and suppress the hyperplasy of the intercapillary cells. All of the actions may be associated with the cytoskeletal protein:nestin and vimentin.