摘要
目的:观察益气清热膏对嘌呤霉素氨基核苷大鼠模型(PAN)的作用机制。方法:40只雄性Wistar大鼠随机分为假手术组、模型组、中药组和环孢素组。通过单次静脉注射PAN(40mg/kg体质量)建立PAN损伤模型。测定血清生化指标和24h尿蛋白定量。观察肾脏病理改变和超微结构变化,RT-PCR及Western Blot检测细胞骨架调控蛋白(α-actinin-4、Synaptopodin、Desmin和uPAR)mRNA和蛋白的定量表达。结果:与模型组比较,益气清热膏可降低PAN大鼠24h尿蛋白定量、改善肾功能(P<0.05);还可减轻肾小球病理损伤,减少足突宽度,改善足突融合,恢复部分足突正常结构;下调肾组织中α-actinin-4、Synaptopodin、Desmin和uPAR的mRNA表达和蛋白表达水平(P<0.05)。结论:益气清热膏能显著改善足细胞损伤,减轻PAN导致的肌动蛋白细胞骨架重组,其作用机制可能与下调肌动蛋白细胞骨架相关调控蛋白表达有关。
Objective: To observe the effect mechanism of Yiqi Qingre Ointment(YQO) on puromycin aminonucleoside rat model(PAN). Methods: Forty male Wistar rats were randomly divided into sham group, model group, YQO group and cyclosporine(CsA) group. A PAN injury model was established by single intravenous injection of PAN(40 mg/kg body weight).Serum biochemical parameters and 24 h urinary protein quantitation was measured on 5 and 9 days after modeling respectively.Renal tissues were harvested for observations of pathologic alterations and ultrastructural changes. Quantitative expressions of regulatory proteins of cytoskeleton were measured using Western Blot and RT-PCR polymerase chain reaction. Result: Compared with the model group, YQO could reduce 24-hour urinary protein and improve renal function in PAN rats(P<0.05), alleviate renal pathological damage, reduce the width of foot processes, attenuate foot process fusion, and restore part of normal structure of foot process. The mRNA and protein expressions of α-actinin-4, Synaptopodin, Desmin and uPAR in kidney tissues were significantly lower than those in kidney tissues(P<0.05). Conclusion: YQO can significantly reduce urinary protein level, repair podocyte injury and reduce actin cytoskeleton reorganization induced by PAN in rats. Its mechanism may be related to the down-regulation of actin cytoskeleton regulatory protein expressions.
引文
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