香椿子正丁醇提取物通过激活Nrf2改善高糖引起的肾小球内皮细胞氧化应激

N-butyl alcohol extract of toona sinensis attenuates the oxidative stress of glomerular endothelial cells induced by high glucose through activating Nrf2

目的探讨香椿子正丁醇提取物(n-butyl alcohol extract of toona sinensis,NBAE)对高糖引起的肾小球内皮细胞氧化应激的影响及机制。方法分别以高糖(high glucose,HG)、HG+NBAE刺激人肾小球内皮细胞,采用DCFDA法检测细胞内活性氧(reactive oxygenspecies,ROS)生成情况,Nitrate/Nitrite Colorimetric法检测细胞内NO的含量,Western blot检测Nrf2、NQO1及HO-1的蛋白表达,免疫荧光方法检测Nrf2、P47phox在细胞内的定位及表达。结果高糖刺激肾小球内皮细胞后出现氧化应激损伤,ROS升高,NO减少,p47phox表达量增高,Nrf2及下游蛋白NQO1、HO-1表达减少;NBAE可明显提高Nrf2的表达,并增加下游蛋白NQO1和HO-1的表达,从而抑制高糖导致的ROS升高,抑制p47phox的表达,并稳定NO的含量。结论NBAE通过激活Nrf2及其下游靶蛋白来改善高糖对肾小球内皮细胞造成的氧化应激损伤。

Objective To investigate the effect of n-butyl alcohol extract of toona sinensis(NBAE)on oxidative stress in glomerular endothelial cells induced by high glucose and its mechanism.Methods Human glomerular endothelial cells were stimulated with high glucose(HG)and HG+NBAE,respectively.Then the production of reactive oxygen species in cells was detected by the DCFDA method.The amount of NO in the cells was measured by the nitrate/nitrite colorimetric method.The expression of Nrf2,NQO1 and HO-1 proteins was detected by Western blot.The localization and expression of Nrf2 and P47phox in the cells were detected by immunofluorescence.Results High glucose induces oxidative stress injury after stimulating the glomerular endothelial cells,and leads to increased ROS,decreased NO,increased expression of p47phox,decreased expression of Nrf2 and its downstream protein NQO1 and HO-1;NBAE can significantly increase the expression of Nrf2 and its downstream protein NQO1 and HO-1,thereby inhibiting high glucose-induced ROS elevation,inhibiting the expression of p47phox,and stabilizing NO content.Conclusion NBAE improves oxidative stress damage induced by high glucose in glomerular endothelial cells by activating Nrf2 and its downstream target proteins.