α-硫辛酸通过mTOR通路抑制缺血再灌注诱导的PC12细胞自噬性凋亡  

α-lipoic acid inhibiting autophagic apoptosis induced by ischemia-reperfusion in PC12 through mTOR pathway

作　　者：谢鹏 任真奎 吕菊 胡玉梅 官志忠[1,2] 张春林 禹文峰[1,2] XIE Peng;REN Zhen-Kui;LV Ju(Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学分子生物学重点实验室,贵州贵阳550004 [2]贵州医科大学地方病与少数民族疾病教育部重点实验室,贵州贵阳550004 [3]黔西南州人民医院检验科 [4]贵州医科大学基础医学院

出　　处：《中国老年学杂志》2021年第3期577-581,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金(81360199);黔科中引地〔2019〕4008号;贵州省卫生健康委科学技术基金(gzwjkj2019-1-039);黔西南州科技局基金(2019-1-10)。

摘　　要：目的研究α-硫辛酸(LA)对PC12细胞缺血再灌注损伤的保护作用及其分子机制。方法传代培养PC12细胞,PC12细胞缺氧/复氧(氧糖剥夺4 h,复氧18 h)模拟缺血再灌注损伤模型。实验分为正常组、缺氧/复氧组、缺氧/复氧+α-LA、缺氧/复氧+自噬抑制剂巴弗洛霉素(Baf)A1组。CCK8筛选α-LA的有效作用浓度;流式细胞术检测细胞凋亡变化;Western印迹检测细胞促凋亡蛋白B细胞淋巴瘤(Bcl-2)相关X蛋白(Bax)、酶切caspase-3、自噬标志分子SQSTM-1/P62、LC3-Ⅱ/Ⅰ及mTOR通路的表达变化。结果缺氧/复氧显著降低PC12细胞的存活率(P<0.05),Western印迹结果显示Bax、酶切caspase-3、LC3-Ⅱ/Ⅰ的表达水平均显著升高(P<0.05),P62和p-mTOR的表达水平降低(P<0.05);与缺氧/复氧组比较,给予α-LA和Baf A1均能显著降低缺氧/复氧诱导的凋亡水平(P<0.05)。结论α-LA通过抑制缺氧/复氧诱导的过度自噬进而减少过度自噬诱导的细胞凋亡。Objective To investigate the protective effect ofα-lipoic acid(LA)on ischemia-reperfusion injury in PC12 cells and its molecular mechanism.Methods PC12 cells were sub-cultured and hypoxia/reoxygenation model(oxygen glucose deprivation for 4 h,reoxygenation for 18 h)was used to simulate the model of ischemia-reperfusion injury in PC12 cells.The experiment was divided into normal group,hypoxia/reoxygenation group,hypoxia/reoxygenation withα-LA group,hypoxia/reoxygenation with Baf.A1 group(autophagy inhibitor Bafilomycin A1).CCK8 screened the effective concentration ofα-LA.Flow cytometry detected the changes of apoptosis.Western blot was used to detect the apoptosis-related protein Bax,Cleaved-caspase-3,autophagy marker molecule P62/SQSTM-1,LC3Ⅱ/Ⅰand mTOR pathway.Results Hypoxia/reoxygenation significantly decreased the survival rate of PC12 cells,the expressions of Bax,Cleaved-caspase-3 and the ratio of LC3Ⅱ/Ⅰwere significantly increased(P<0.05),the expressions of P62 and p-mTOR were decreased significantly(P<0.05),andα-LA and Baf.A1 could significantly decrease the apoptosis level induced by hypoxia/reoxygenation compared with hypoxia/reoxygenation group(P<0.05).Conclusionsα-LA could reduce apoptosis induced by hypoxia/reoxygenation by inhibiting excessive autophagy.Therefore,this study might provide reference for clinical drug use or auxiliary drug use for ischemic stroke.

关 键 词：Α-硫辛酸 缺血再灌注损伤 自噬 自噬性凋亡 

分 类 号：R965.1[医药卫生—药理学] R743[医药卫生—药学]