基于Nrf2/HO-1通路探讨黄芪-当归药对改善糖尿病小鼠肾脏氧化损伤的机制

Mechanism research of Astragalus-Angelica medicine pairs on improving renal oxidative damage in diabetic mice based on Nrf2/HO-1 pathway

目的:探讨不同配比的黄芪-当归(芪归)药对对糖尿病(DM)小鼠肾脏的保护作用及分子机制。方法:选取雄性C57BL/6小鼠80只,采用高脂高糖饮食联合链脲佐菌素(STZ)腹腔注射的方法制备2型糖尿病小鼠模型,成模后随机分为模型组、不同配比芪归(芪归5∶1、2∶1、1∶1、1∶2、1∶5)组、缬沙坦组;同周龄C57BL/6小鼠10只作为对照组。治疗组予相应浓度的药物灌胃,模型组、对照组予等体积0.9%氯化钠溶液。12周后处死小鼠,检测各组小鼠肾功能、尿ACR及MDA、T-AOC、T-SOD、CAT、GSH-PX水平,RT-PCR、Western Blot法分别检测小鼠肾脏Nrf2、HO-1 mRNA及蛋白表达水平,同时应用免疫组化法检测Nrf2水平。结果:与对照组比较,模型组小鼠血清Cys-C、β2-MG,尿ACR,肾脏MDA,Nrf2、HO-1 mRNA表达水平显著升高(P<0.01),T-AOC及T-SOD、CAT、GSH-PX表达显著降低(P<0.01,P<0.05);与模型组比较,不同配比芪归药对能够不同程度减少小鼠尿ACR、改善Cys-C、β2-MG、减轻肾脏组织MDA、T-AOC水平,增加小鼠肾组织Nrf2、HO-1mRNA的表达(P<0.01,P<0.05)。结论:芪归药对能够改善DM小鼠肾脏损伤,可能与激活Nrf2/HO-1通路,增强抗氧化反应有关。

Objective:To investigate the protective effect and molecular mechanism of Astragalus-Angelica medicine pairs at different proportions on kidney of diabetic mice.Methods:A total of 80 male C57BL/6 mice were selected and used high-fat and high-sugar diet combined STZ intraperitoneal injection to prepare type 2 diabetic mouse models.After modeling,they were randomly divided into model groups,Astragalus-Angelica medicine pairs(Astragalus-Angelica 5∶1,2∶1,1∶1,1∶2,1∶5)groups and valsartan group;10 C57BL/6 mice of the same age as the normal group.The treatment groups were given the corresponding concentration of drugs by gavage,and the model group and the normal group were given equal volume of normal saline.After 12 weeks,the mice were sacrificed and the renal function,urine ACR,MDA,T-AOC,T-SOD,CAT and GSH-PX levels were detected in each group.RT-PCR and Western Blot were used to detect the expression levels of Nrf2,HO-1 mRNA and protein in the kidney of the mice,and the Nrf2 level was detected by immunohistochemistry.Results:Compared with the normal group,the expression levels of serum Cys-C,β2-MG,urine ACR,kidney MDA,Nrf2,HO-1 mRNA in the model group were significantly increased(P<0.01),while T-AOC and the contents of T-SOD,CAT,GSH-PX were significantly decreased(P<0.01,P<0.05).Compared with the model group,different ratios of Astragalus-Angelica medicine pairs could reduce ACR,improve Cys-C,β2-MG,reduce kidney tissue MDA,T-AOC level,and increase the expression of Nrf2,HO-1 mRNA in kidney tissue.Conclusion:Astragalus-Angelica medicine pairs can improve the kidney damage of diabetic mice,which may be related to the activation of Nrf2/HO-1 pathway and the enhancement of antioxidant response.