白虎加人参汤对高尿酸血症肾病小鼠保护作用及机制研究

Protective effect and mechanism of Baihu Jia Renshen Decoction on mice with hyperuricemic nephropathy

目的白虎加人参汤对高尿酸血症肾病(HN)小鼠的保护作用及机制研究。方法将C57BL/6小鼠48只随机分为正常组、模型组、阳性药物组和白虎加人参汤组。正常组小鼠灌胃0.5%羧甲基纤维素钠(CMC-Na),其余各组小鼠灌胃次黄嘌呤500 mg/kg和腹腔注射氧嗪酸钾200 mg/kg诱导高尿酸血症肾病小鼠模型,阳性药物组每日灌胃非布司他5 mg/kg,白虎加人参汤组小鼠每日灌胃白虎加人参汤54 g/kg,连续给药21d。采集小鼠血清及肾脏组织,测定小鼠血清中SUA、Scr和BUN含量;HE染色,镜下观察小鼠肾脏的病理变化;测定小鼠肾脏组织中GSH-Px、SOD、CAT及MDA水平;ELISA法测定小鼠血清中TNF-α、IL-6和IL-1β表达;Western blot法测定小鼠肾脏组织中AKT/GSK-3β/Nrf2信号通路和NLRP3炎性小体相关蛋白表达。结果生化结果显示,白虎加人参汤能够降低高尿酸血症肾病小鼠血清中SUA、Scr和BUN水平。HE染色结果显示,白虎加人参汤可以减轻高尿酸血症肾病小鼠肾脏损伤。白虎加人参汤能够抑制高尿酸血症肾病小鼠肾脏组织中MDA表达,促进GSHPx、SOD和CAT的表达,抑制TNF-α、IL-6和IL-1β表达;Western blot结果显示,白虎加人参汤能够促进高尿酸血症肾病小鼠肾脏组织中p-AKT、p-GSK-3β、Nrf2和HO-1蛋白表达,抑制NLRP3、Caspase1、IL-1β蛋白表达。结论白虎加人参汤通过激活AKT/GSK-3β/Nrf2信号通路,抑制NLRP3炎性小体,改善高尿酸血症肾病小鼠氧化应激和炎症,减轻高尿酸血症肾病。

Objective To study the protective effect and mechanism of Baihu Jia Renshen Decoction(BRD)on mice with hyperuricemic nephropathy(HN).Methods Forty-eight C57BL/6 mice were randomly divided into four groups:a normal group,a model group(HN),a positive drug(febuxostat,HN+FBX)group,and a Baihu Jia Renshen Decoction(HN+BRD)group.The mice in the normal group were given 0.5%carboxymethyl cellulose sodium(CMC-Na)by gavage,while the mice in the other groups were given 500 mg/kg hypoxanthine by gavage and 200 mg/kg potassium oxyzinate by intraperitoneal injection to induce HN models.The mice in the HN+FBX group and the HN+BRD group were,respectively,given 5 mg/kg FBX and 54 g/kg BRD by gavage daily,both for 21 days.Their serum and kidney tissue were collected,and the serum levels of SUA,Scr and BUN were measured.HE staining was used to observe the pathological changes of the kidney under microscope.The levels of GSH-Px,SOD,CAT and MDA in kidney tissue were measured.ELISA was used to measure the expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in serum.Western blot was used to measure the protein expression of AKT/GSK-3β/Nrf2 signaling pathway and NLRP3 inflammasome in kidney tissue of mice.Results The biochemical results showed that BRD could effectively reduce the levels of SUA,Scr and BUN in the serum of HN mice.The HE results showed that BRD could improve the kidney injury of HN mice.BRD could inhibit the expression of MDA,promote the expression of GSH-Px,SOD and CAT in kidney tissue of HN mice,inhibit the expression of TNF-α,IL-6 and IL-1βin the serum of HN mice.The Western blot results showed that BRD could promote the protein expression of p-AKT,p-GSK-3β,Nrf2 and HO-1 in the kidney tissue of HN mice,and inhibit the protein expression of NLRP3,Caspase1 and IL-1βin the kidney tissue of HN mice.Conclusion BRD activates the AKT/GSK-3β/Nrf2 signaling pathway and inhibits NLRP3 inflammasome,improving oxidative stress and inflammation in HN mice,thereby reducing hyperuricemic nephropathy in mice.