四妙散改善高尿酸血症和肾损伤以及调节肾脏线粒体相关蛋白的作用研究

Effects of Simiao San on alleviation of hyperuricemia and renal injury and regulation of renal mitochondria related proteins

目的探索四妙散改善高尿酸血症小鼠尿酸代谢和肾损伤的作用和可能的作用机制。方法采用氧嗪酸钾(300 mg/kg)联合次黄嘌呤(500 mg/kg)建立急性高尿酸血症小鼠模型,然后给予四妙散水提液(4.55、9.1、18.2 g/kg)干预7 d。利用生化法分别测定血清和尿液中尿酸(uric acid,UA)的含量,血清中超氧化物歧化酶(superoxide dismutase,SOD)活力及氧化型谷胱甘肽(oxidized glutathione,GSSG)、丙二醛(malondialdehyde,MDA)和肾脏中还原型谷胱甘肽(reduced glutathione,GSH)含量,以及肝脏中黄嘌呤氧化酶(xanthine oxidase,XOD)的活力。利用Western blotting法检测肾脏线粒体融合和分裂相关蛋白表达。苏木素-伊红(HE)染色法观察肾脏的病理变化。结果四妙散显著降低高尿酸血症小鼠血清中UA含量(P<0.05、0.01),显著升高尿液中UA含量(P<0.05、0.001),显著升高血清SOD活力和肾脏中GSH水平(P<0.05、0.01、0.001),显著降低血清GSSG和MDA水平(P<0.05、0.001),显著降低肝脏中XOD活性(P<0.05)。同时,四妙散可显著升高肾脏线粒体融合蛋白1(mitofusin 1,MFN1)、MFN2、视神经萎缩相关蛋白1(optic atrophy 1,OPA1)、过氧化物酶体增殖物活化受体γ辅助活化因子-1α(peroxisome proliferator-activated receptor gamma coactivator-1α,PGC-1α)蛋白水平(P<0.05),降低线粒体动力相关蛋白1(dynamin-related protein1,DRP1)蛋白表达水平(P<0.05),进而抑制其肾脏的病理改变。结论四妙散能通过调节线粒体相关蛋白的表达,增强机体抗氧化应激的能力,进而改善尿酸代谢和抑制肾损伤。

Objective To explore the effect and possible mechanism of Simiao San(四妙散)on improving uric acid metabolism and kidney damage in hyperuricemia mice.Methods The acute hyperuricemia mouse model was established with oteracil potassium(300 mg/kg)and hypoxanthine(500 mg/kg),and then the water extract of Simiao San(4.55,9.1,18.2 g/kg)was given for 7 d.The content of uric acid(UA)in serum and urine,the activities of superoxide dismutase(SOD)and contents of oxidized glutathione(GSSG),malondialdehyde(MDA)in serum,and reduced glutathione(GSH)level in kidney,and activity of xanthine oxidase(XOD)in liver were measured by biochemical method.Western blotting was used to detect the expressions of fusion and division related proteins in kidney mitochondria.The pathological changes of kidney was observed by HE staining.Results Simiao San significantly reduced UA content in serum of hyperuricemia mice(P<0.05,0.01),significantly increased UA content in urine(P<0.05,0.001),significantly increased SOD activity in serum and GSH level in kidney(P<0.05,0.01,0.001),significantly reduced GSSG and MDA levels in serum(P<0.05,0.001),and significantly reduced XOD activity in liver(P<0.05).At the same time,Simiao San significantly increased renal mitofusin protein 1(MFN1),MFN2,optic atrophy 1(OPA1)and peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)protein expressions(P<0.05),reduced dynamin-related protein 1(DRP1)protein expression(P<0.05),thereby inhibiting the pathological changes of the kidney.Conclusion Simiao San can enhance ability of body to resist oxidative stress by regulating the expression of mitochondrial related proteins,thereby improving uric acid metabolism and inhibiting kidney damage.