摘要
目的探讨早期跑步通过热休克蛋白A5(HSPA5)调节PINK1/Parkin介导的线粒体自噬改善脑缺血/再灌注(I/R)小鼠的记忆功能。方法将雄性BALB-c小鼠随机分为Sham组、跑步组、I/R组、跑步+I/R组、跑步+I/R+3-MA(自噬抑制剂)组、跑步+I/R+scramble siRNA组和跑步+I/R+HSPA5 siRNA组,每组8只。I/R 7 d后,采用Longa评分法评估各组小鼠神经功能;TTC染色评估各组小鼠脑梗死体积;水迷宫实验分析评估各组小鼠记忆功能;ELISA检测各组小鼠血清肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β和IL-10含量;实时定量PCR检测各组小鼠缺血半影区HSPA5 mRNA水平;Western blotting检测各组小鼠缺血半影区PINK1和Parkin蛋白表达。结果与Sham组相比,I/R组小鼠神经功能缺损评分和脑梗死体积增加;目标象限停留时间和穿越平台次数减少;血清TNF-α和IL-1β水平升高,而IL-10水平降低(P均<0.05)。与I/R组相比,跑步+I/R组小鼠神经功能缺损评分和脑梗死体积均降低;目标象限停留时间和穿越平台次数增加;血清TNF-α和IL-1β水平下降,同时IL-10水平上升(P均<0.05)。与跑步+I/R组相比,跑步+I/R+HSPA5 siRNA组小鼠目标象限停留时间和穿越平台次数减少;血清TNF-α和IL-1β水平升高,而IL-10水平降低(P均<0.05)。与Sham组相比,I/R组小鼠缺血半影区HSPA5 mRNA及PINK1、Parkin蛋白表达增加(P均<0.05);与I/R组相比,跑步+I/R组小鼠HSPA5 mRNA及PINK1、Parkin蛋白表达进一步增加(P均<0.05);与跑步+I/R组相比,跑步+I/R+3-MA组和跑步+I/R+HSPA5 siRNA组小鼠PINK1和Parkin蛋白表达均降低(P均<0.05)。结论早期跑步运动可能通过上调HSPA5并激活PINK1/Parkin介导的线粒体自噬改善I/R小鼠的记忆功能。
Objective To explore how early running improves memory function in mice with cerebral ischemia/reperfusion(I/R)injury by modulating PINK1/Parkin-mediated mitophagy through heat shock protein A5(HSPA5).Methods Male BALB/c mice were randomly divided into the sham,running,I/R,running+I/R,running+I/R+3-MA(autophagy inhibitor),running+I/R+scrambled siRNA,and running+I/R+HSPA5 siRNA groups,with eight mice in each group.After seven days of I/R,the neurological function of mice in each group was evaluated by the Longa score,cerebral infarct volume by TTC staining,memory function by water maze analysis,serum levels of tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β)and interleukin-10(IL-10)by ELISA,mRNA level of HSPA5 in the ischemic penumbra of mice by real time quantitative PCR,and protein expressions of PINK1 and Parkin in the ischemic penumbra by Western blotting.Results Compared with the sham group,the I/R group had increased neurological deficit score and infarct volume,decreased time spent in the target quadrant and number of platform crossings,increased serum levels of TNF-αand IL-1β,and decreased level of IL-10(all P<0.05).Compared with the I/R group,the running+I/R group had decreased neurological deficit score and infarct volume,increased time spent in the target quadrant and the number of platform crossings,decreased serum levels of TNF-αand IL-1β,and increased level of IL-10(all P<0.05).Compared with the running+I/R group,the running+I/R+HSPA5 siRNA group had decreased time spent in the target quadrant and the number of platform crossings,increased serum levels of TNF-αand IL-1β,and decreased level of IL-10(all P<0.05).Compared with the sham group,the I/R group had increased expression of HSPA5 mRNA and PINK1 and Parkin protein in the ischemic penumbra(all P<0.05).The expression of HSPA5 mRNA and PINK1 and Parkin protein was further increased in the running+I/R group compared with the I/R group(all P<0.05),while the expression of PINK1 and Parkin protein was decreased in the running+I/R+3-MA and the running+I/R+HSPA5 siRNA groups compared with the running+I/R group(all P<0.05).Conclusion Our study provided evidence showing that early running could improve memory function in mice with I/R injury by activation of PINK1/Parkin-mediated mitophagy through the upregulation of HSPA5.
作者
彭志锋
李加善
张继红
PENG Zhifeng;LI Jiashan;ZHANG Jihong(Department of Physiology,School of Medicine,Shanxi Datong University,Datong 037009,China;Department of Anatomy,School of Medicine,Shanxi Datong University,Datong 037009,China)
出处
《中国医科大学学报》
北大核心
2026年第2期122-126,132,共6页
Journal of China Medical University
基金
山西省科技厅基础研究项目(202303021211326)。
