益气清热膏通过肌动蛋白骨架重排保护嘌呤霉素氨基核苷大鼠足细胞损伤机制

Protective mechanism of Yiqi Qingre Ointment on podocyte injury in rat model of purinomycin aminonucleoside by actin skeleton rearrangement

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 ofα-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.