青蒿琥酯通过激活Nrf2通路增强小鼠脑梗死后的抗氧化能力和减轻神经损伤

Artesunate improved antioxidant defenses and reduced brain damage by activiting Nrf2 pathway in a mouse stroke model

目的探讨青蒿琥酯是否能减轻脑梗死小鼠脑损伤和增强抗氧化作用,以及其对核因子E2相关因子2(Nrf2)抗氧化通路的影响。方法将小鼠分为4组:dMCAO组、假手术组、青蒿琥酯低剂量组(15mg·kg^(-1))和青蒿琥酯高剂量组(30mg·kg^(-1))。脑缺血后72h,通过TTC染色测量的梗死体积和神经功能评分来评价青蒿琥酯的神经保护作用。用试剂盒分别测定缺血侧脑组织中MDA、ROS、SOD、CAT和GSH-Px的含量及活性。Western blot检测缺血脑组织中血红素氧合酶-1(HO-1)和Nrf2的蛋白水平。免疫荧光分析观察Nrf2的核转位。结果与dMCAO组相比,青蒿琥酯30mg·kg^(-1)对梗死体积和神经功能缺损有明显的改善作用。青蒿琥酯30mg·kg^(-1)处理可降低MDA和ROS的产生,提高CAT、SOD和GSHPx活性,同时能显著提高HO-1的蛋白水平,并上调Nrf2在细胞核中的表达。结论青蒿琥酯增强了脑缺血后的抗氧化作用,保护脑组织,其作用可能通过Nrf2信号通路介导。

Objective The purpose of our experiment was to investigate whether artesunate attenuated brain damage and activated antioxidant defense.Also,we investigated the influnence of artesunate in the Nuclear factor-E2-related factor 2(Nrf2)pathway.Methods The mice were divided into four groups:dMCAO group,sham group,low dose group(artesunate treated 15mg·kg^(-1))and high dose group(artesunate treated 30mg·kg^(-1)).We analyzed the neuroprotective effect of artesunate at 72 hours after cerebral ischemia,through evaluation infarct volume by TTC stain and assessment neurological score.In ischemic brain tissue,MDA,ROS,SOD,CAT and GSH-Px were detected by kit respectively.The protein levels of HO-1and Nrf2 were confirmed by Western blot.The nuclear translocating of Nrf2 was detected by Western blot and immunofluorescence.Results It showed that artesunate 30mg·kg^(-1)had improving effect on the infarct volume and neurological deficit scores,compared to dMCAO group.Treatment with artesunate 30mg·kg^(-1)resulted in the decrease of MDA and ROS and the increase of CAT,SOD and GSH-Px activities.Meantime artesunate upregulated the protein levels of HO-1 and Nrf2 significantly.Furthermore,Artesunate 30mg·kg^(-1)upregulated the expression level of Nrf2 in nuclear.Conclusion In conclusion,artesunate improved antioxidant defenses and protect brain after cerebral ischemia and the effect might be mediated by Nrf2 signaling pathway,which downstream p rotein can protect brain against oxidative stress.