自噬在氧糖剥夺诱导星形胶质细胞损伤中的作用机制  

Mechanism of autophagy in astrocyte damage induced by oxygen-glucose deprivation

作　　者：秦爱萍 王锦淳 王蕾 周杰 冯丁丁 QIN Aiping;WANG Jinchun;WANG Lei(School of Pharmacy,Jiangsu Health Vocational College,Nanjing 211800,CHINA)

机构地区：[1]江苏卫生健康职业学院药学院,江苏南京211800

出　　处：《江苏医药》2022年第12期1192-1195,F0002,共5页Jiangsu Medical Journal

基　　金：江苏高校“青蓝工程”优秀青年骨干教师项目(苏教师函[2022]29号);江苏省卫生健康委科研项目(M2021057)。

摘　　要：目的 探讨自噬在氧糖剥夺(OGD)诱导星形胶质细胞损伤中的作用机制。方法 培养大鼠原代星形胶质细胞,分为对照组和OGD组。OGD组体外建立OGD损伤模型,OGD作用0.5、1、3、6和12 h,采用免疫荧光鉴定原代星形胶质细胞纯度,Western blot法检测微管相关蛋白1A/1B-轻链3(LC3)蛋白表达。另设OGD+3-甲基腺嘌呤(3-MA)组,予自噬抑制剂3-MA 10 mmol/L,OGD作用3 h。免疫荧光检测细胞中LC3蛋白表达,乳酸脱氢酶(LDH)法检测细胞损伤程度。结果 原代星形胶质细胞纯度可达95%以上。与对照组比较,OGD作用1、3 h时LC3-Ⅱ/LC3-Ⅰ比值升高,且作用3 h时LC3-Ⅱ/LC3-Ⅰ比值更高(P<0.01)。OGD组可见较多LC3绿色荧光散在分布于细胞质内,OGD+3-MA组细胞质内LC3荧光强度减弱。OGD组LDH漏出率高于对照组(P<0.01),OGD+3-MA组LDH漏出率低于OGD组(P<0.05)。结论 自噬参与并促进OGD诱导星形胶质细胞损伤。Objective To explore the mechanism of autophagy in astrocyte damage induced by oxygen-glucose deprivation(OGD).Methods Primary astrocytes of rats were cultured and divided into two groups of OGD and control. OGD damage model was established in vitro acted with OGD for 0.5, 1, 3, 6 and 12 hours. Purity of primary astrocytes was identified by immunofluorescence, and microtubule-associated protein 1A/1B light chain 3(LC3) protein was detected by Western blot. Another OGD+3-MA group was set up to give autophagy inhibitor 3-MA 10 mmol/L, and OGD was applied for 3 hours. LC3 protein and degree of cell damage were detected by immunofluorescence and LDH method, respectively.Results The purity of primary astrocytes reached at 95% or more. Compared with control group, the ratio of LC3-Ⅱ to LC3-Ⅰ was increased at the times of acted with OGD for 1 and 3 hours, and which was higher at OGD 3 hours(P<0.01). More LC3 green fluorescence was scattered in the cytoplasm of OGD group, and the fluorescence intensity of LC3 in the cytoplasm of OGD+3-MA group was weakened. The LDH leakage rate in OGD group was higher than that in control group(P<0.01),and the LDH leakage rate in OGD+3-MA group was lower than that in OGD group(P<0.05).Conclusion Autophagy participates in and promotes OGD-induced astrocyte damage.

关 键 词：氧糖剥夺 星形胶质细胞 自噬 微管相关蛋白1A/1B-轻链3 

分 类 号：R965[医药卫生—药理学]