贵州医科大学学报

2019, v.44;No.222(03) 259-263

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尼古丁通过PI3K/AKT信号通路上调星形胶质细胞HSF1的表达
Nicotine Upregulates HSF1 Expression via PI3K/AKT Signaling Pathway in Astrocytes

董智慧;任真奎;吕菊;谢鹏;官志忠;吴昌学;禹文峰;
DONG Zhihui;REN Zhenkui;LV Ju;XIE Peng;GUAN Zhizhong;WU Changxue;YU Wenfeng;Key Laboratory of Medical Molecular Biology,Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University;Department of Clinical Laboratory,People's Hospital,Qianxinan Buyei and Miao Autonomous Prefecture;Department of Pathology,Guizhou Medical University;

摘要(Abstract):

目的:研究尼古丁对星形胶质细胞热休克转录因子1(HSF1)的表达的影响,并探讨其机制。方法:分离新出生24 h内乳鼠大脑皮质,进行原代培养并鉴定为星形胶质细胞,待细胞成熟后分为正常对照组、尼古丁处理组,PI3K/AKT信号通路阻断剂LY294002处理组和尼古丁+LY294002处理组,正常对照组不做任何处理;尼古丁处理组用5μmol/L尼古丁分别处理星形胶质细胞6、12、18、24 h; LY294002处理组只加10μmol/L LY294002处理;尼古丁+LY294002处理组是先加10μmol/L LY294002处理2 h后,再加入5μmol/L尼古丁共同处理18 h,免疫印迹法观察各组HSF1蛋白的表达。结果:与正常对照组相比,尼古丁处理组星形胶质细胞内HSF 1蛋白的表达上调(P <0. 05);与尼古丁处理组比较,LY294002+尼古丁处理组星形胶质细胞内HSF1表达明显受到抑制(P <0. 05)。结论:尼古丁通过PI3K/AKT信号通路上调星形胶质细胞HSF1蛋白表达。
Objective: To study the effect of nicotine on the expression of heat shock transcription factor 1( HSF1) in astrocytes and explore its mechanism. Methods: To get primary rat astrocytes,the cerebral cortexes of newborn rats within 24 h were isolated,prepared for single cell suspension,cultured and verified using GFAP immunostaining. After astrocytes matured,they were divided into four groups: control group,nicotine group,LY294002( PI3 K/AKT inhibitor) group and the nicotine +LY294002 group. The control group was not given any treatment,; the nicotine group was treated with5 μmol/L nicotine for 6 h,12 h,18 h and 24 h,respectively; LY294002 group was given 10 μmol/L LY294002; nicotine + LY294002 group was pretreated with 10 μmol/L LY294002 for 2 h and then treated with 5 μmol/L nicotine for 18 h. The expression levels of HSF1 protein were detected by immunoblotting. Results: Compared with the control group,HSF-1 level was upregulated in nicotine group( P < 0. 05). HSF1 expression level was remarkably lower in LY294002 + nicotine group than that in nicotine group( P < 0. 01). Conclusion: Nicotine upregulates HSF1 expression through PI3 K/AKT signaling pathway in astrocytes.

关键词(KeyWords): 原代星形胶质细胞;热休克核转录因子1;尼古丁;PI3K/AKT信号通路;阿尔茨海默症
primary astrocytes;heat shock nuclear transcription factor 1;nicotine;PI3K/AKT pathway;Alzheimer's disease

Abstract:

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基金项目(Foundation): 国家自然科学基金(批准号:81360199);; 教育部科学技术研究项目(213032A);; 贵州省国际科技合作计划项目[黔科合外G字(2013)7026号];; 贵州省创新计划项目[黔教合协同创新中心(2014)06];; 贵州省教育厅项目(2015年贵州省普通高等学校地方病和少数民族疾病防控创新团队);贵州省教育厅项目[黔教合外G字(2013)63];; 贵州省科技厅计划课题[黔科合重大专项字(2014)6008]

作者(Author): 董智慧;任真奎;吕菊;谢鹏;官志忠;吴昌学;禹文峰;
DONG Zhihui;REN Zhenkui;LV Ju;XIE Peng;GUAN Zhizhong;WU Changxue;YU Wenfeng;Key Laboratory of Medical Molecular Biology,Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University;Department of Clinical Laboratory,People's Hospital,Qianxinan Buyei and Miao Autonomous Prefecture;Department of Pathology,Guizhou Medical University;

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