【摘要】 目的探讨β-细辛醚对淀粉样前体蛋白(APP)/早老素1(PS1)小鼠的作用机制及对PINK1/Parkin介导的线粒体自噬的影响。方法将APP/PS1小鼠随机分为模型组、β-细辛醚低、中、高剂量组(15、30、45 mg/kg)、多奈哌齐组(1 mg/kg)、雷帕霉素组(4 mg/kg)、3-甲基腺嘌呤(3-MA)组(4 mg/kg)、环孢霉素A组(10 mg/kg),另设正常组,每组10只。各组灌胃给药,模型组和正常组用等体积蒸馏水代替,1次/d,给药30 d。观察各组小鼠水迷宫实验,蛋白质印迹法检测小鼠海马中APP、PS1、神经突触素1(SYN1)、β-位点淀粉样前体蛋白裂解酶1(BACE1)、LC3 I/II、Beclin-1、p62、PINK1、Parkin和p-Parkin蛋白表达水平;反转录聚合酶链反应检测APP、PS1、BACE1、LC3B、Beclin-1、p62和Parkin mRNA表达水平。结果与模型组相比,β-细辛醚高剂量组、β-细辛醚中剂量组和多奈哌齐组APP/PS1小鼠的逃避潜伏期缩短(P<0.05或P<0.01),小鼠海马内APP、PS1、BACE1的表达下降(P<0.01),SYN1表达上升(P<0.05或P<0.01);与模型组相比,β-细辛醚中剂量组APP/PS1小鼠内Beclin-1、LC3B表达均升高(P<0.05或P<0.01),p62表达下降(P<0.05);与模型组相比,雷帕霉素组Beclin-1、LC3B表达均升高(P<0.05或P<0.01),p62表达下降(P<0.01);与模型组相比,3-MA组p62表达升高(P<0.05);与模型组相比,β-细辛醚中剂量组APP/PS1小鼠内PINK1、p-Parkin表达均显著升高(P<0.01)。结论β-细辛醚通过激活PINK1-Parkin介导的线粒体自噬而影响阿尔茨海默病病理,这为应用β-细辛醚治疗阿尔茨海默病提供了新的靶点。更多还原

【Abstract】 Objective To explore the mechanism of β-asarone on APP/PS1 mice and its effect on the mitochondrial autophagy mediated by PINK1/Parkin. Methods APP/PS1 mice were randomly divided into model group, low/medium/high dose groups(15, 30, 45 mg/kg) of β-asarone, donepezil group(1 mg/kg), rapamycin group(4 mg/kg), 3-methyladenine(3-MA) group(4 mg/kg),cyclosporine A(CsA) group(10 mg/kg) and normal group, with 10 mice in each group. Each group was given intragastric administration, and model group and normal group were given equal volume distilled water once a day for 30 days. Water maze test was observed in each group. Amyloid precursor protein(APP), prosenoxin 1(PS1), synaptophysin 1(SYN1), β-site amyloid precursor protein lyase 1(BACE1) and LC3 I/II, Beclin-1, p62, PINK1, Parkin and p-Parkin were detected in the hippocampus of mice by western blot. The mRNA expression levels of APP, PS1, BACE1, LC3 B, Beclin-1, P62 and Parkin were detected by reverse transcription polymerase chain reaction. Results Compared with the model group, the escape latency of APP/PS1 mice inβ-asarone high dose group, β-asarone medium dose group and donepezil group was significantly shortened(P<0.05 or P<0.01), the expression of APP, PS1 and BACE1 in hippocampus of mice decreased(P<0.01), and the expression of SYN1 increased(P<0.05 or P<0.01); compared with the model group, the expression of Beclin-1 and LC3 B in β-asarone medium dose group increased(P<0.01 or P<0.05), and the expression of p62 decreased(P<0.05); compared with the model group, the expression of Beclin-1 and LC3 B in rapamycin group increased(P<0.05 or P<0.01), and the expression of p62 decreased(P<0.01); the expression of p62 in 3-MA group was higher than that model group(P<0.05); compared with the model group, the expression levels of PINK1 and p-Parkin in APP/PS1 mice in β-asarone medium dose group were significantly increased(P<0.01). Conclusion β-asarone may affect Alzheimer’s Disease(AD) pathology by activating mitochondrial autophagy mediated by PINK1-Parkin, which provides a new target for the treatment of AD with β-asarone.更多还原