益气活血通络方对单侧输尿管梗阻大鼠肾脏线粒体动力学的影响

Effect of Yiqi Huoxue Tongluo Recipe on Renal Mitochondrial Dynamics in Rats with Unilateral Ureteral Obstruction

目的 探讨益气活血通络方通过调节线粒体动力学改善肾脏损伤的作用机制。方法 将健康雄性Wistar大鼠48只随机分为假手术组、模型组、氯沙坦组、益气活血通络方组,每组12只。采用单侧输尿管结扎手术复制肾纤维化模型,灌胃给药,连续干预14 d。采用HE和Masson染色法观察肾组织的病理形态变化;免疫组化法检测单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾组织中线粒体融合蛋白(mitofusin,Mfn)2及动力相关蛋白(dynamin-relatedprotein,Drp)1的表达;Western blot检测Mfn2、Drp1、半胱氨酸天冬氨酸蛋白酶(cysteinyl aspartate specific proteinase,Caspase)-3及B细胞淋巴瘤(B-cell lymphoma,Bcl)-2的表达;免疫荧光检测Ⅰ型胶原(Collagen Ⅰ)的表达。结果 与假手术组比较,模型组大鼠肾小管管腔扩张明显,小管萎缩,上皮细胞坏死、脱落,Collagen Ⅰ、Caspase-3及Drp1表达均明显升高(P<0.05),Mfn2及Bcl-2表达均明显降低(P<0.05);与模型组比较,益气活血通络方和氯沙坦组大鼠肾小管上皮细胞萎缩变性及肾间质纤维化程度显著改善,Collagen Ⅰ、Caspase-3及Drp1表达均明显降低(P<0.05),Mfn2及Bcl-2表达均明显升高(P<0.05)。结论 益气活血通络方可调节UUO大鼠肾脏线粒体动力学,改善肾脏病理损伤。

Objective To explore the mechanism of Yiqi Huoxue Tongluo Recipe in improving renal injury by regulating mitochondrial dynamics.Methods 48 male Wistar rats were randomly divided into sham-operated group,model group,losartan group,and Yiqi Huoxue Tongluo Recipe group,with 12 rats in each group.The unilateral ureteral obstruction surgery was used to replicate the renal fibrosis model,and the drug was administered by gavage for 14 consecutive days.Pathomorphological changes in renal tissue was detected by HE staining and Masson staining,the expression of Mfn2 and Drp1 in renal tissues of rats with unilateral ureteral obstruction(UUO)were detected by immunohistochemical staining,the expression of Mfn2,Drp1,Caspase-3 and Bcl-2 were detected by Western blot,the expression of Collagen I was detected by immunofluorescence.Results Compared with the sham-operated group,rats in the model group showed significant tubular lumen dilatation,tubular atrophy,epithelial cell necrosis and detachment,significantly higher expression of Collagen I,Caspase-3 and Drp1(P<0.05),significantly lower expression of Mfn2 and Bcl-2(P<0.05).Compared with the model group,the expression of Collagen I,Caspase-3 and Drp1 were significantly reduced(P<0.05),and the expression of Mfn2 and Bcl-2 were significantly increased(P<0.05)in rats of Yiqi Huoxue Tongluo Recipe group and losartan group.Conclusion The Yiqi Huoxue Tongluo Recipe can regulate renal mitochondrial dynamics in UUO rats and improve their renal pathological injury.