高硒对L02细胞胰岛素信号通路PI3K-AKT-mTOR的影响  

Effect of high selenium on insulin signaling pathway PI3K-AKT-mTOR in L02 cells

作　　者：王琴 张雪 王建荣 刘轶群 韩枫 向雪松 郭岩彬[2] 黄振武 Wang Qin;Zhang Xue;Wang Jianrong;Liu Yiqun;Han Feng;Xiang Xuesong;Guo Yanbin;Huang Zhenwu(National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China;College of Resources and Environmental Sciences,China Agricultural University,Beijing 100193,China;The Key Laboratory of Trace Element Nutrition,National Health Commission of the People's Republic of China,Beijing 100050,China)

机构地区：[1]中国疾病预防控制中心营养与健康所,北京100050 [2]中国农业大学资源与环境科学学院,北京100193 [3]国家卫生健康委微量元素与营养重点实验室,北京100050

出　　处：《卫生研究》2024年第1期77-80,87,共5页Journal of Hygiene Research

基　　金：国家自然科学基金面上项目(No.81973048)。

摘　　要：目的观察高硒对L02细胞胰岛素信号通路PI3K-AKT-mTOR的影响。方法在普通培养基中添加硒代蛋氨酸,使细胞培养液中硒的终浓度分别为0.001、0.0025、0.005、0.0075、0.01、0.025、0.05、0.075和0.1μmol/L,生理盐水为对照组。利用这些高硒培养基培养L02细胞,干预48 h后,一半细胞用PBS清洗,更换为无血清培养基,饥饿4 h后用1μmol/L胰岛素刺激15 min,收集细胞进行裂解,免疫印迹法分析L02细胞裂解液中胰岛素信号通路磷脂酰肌醇3激酶(PI3K)-蛋白激酶B(AKT)-雷帕霉素靶蛋白(mTOR)信号转导通路相关酶的表达水平。另一半细胞在干预48 h后即收集细胞上清和裂解液,免疫印迹法分析细胞裂解液中的谷胱甘肽过氧化物酶1(glutathione peroxidase 1,GPX1)和上清中的硒蛋白P(SELENOP)表达水平。结果高硒培养环境中L02细胞内PI3K-AKT-mTOR信号通路关键酶磷酸化蛋白激酶B(Ser-473)[P-AKT-(Ser-473)]、磷酸化蛋白激酶B(Thr-308)[P-AKT-(Thr-308)]、PI3K和mTOR蛋白表达均随着硒代蛋氨酸浓度的升高而下调。硒蛋白GPX1和SELENOP随着培养基中硒含量升高而表达增强。结论高硒培养的L02细胞内胰岛素信号通路PI3K-AKT-mTOR出现受损。OBJECTIVE To observe the effect of high selenium on insulin signaling pathway PI3K-AKT-mTOR in LO2 cells.METHODSOne group of L02 cell was treated with different concentrations of selenomethionine(SeMet,0.001,0.0025,0.005,0.0075,0.01,0.025,0.05,0.075 and 0.1μmol/L)for 48 h,then cultured with serum-free medium for 4 h and stimulated with 1μmol/L insulin for 15 min.The insulin signaling pathway(PI3K-AKT-mTOR)was detected by WB.Another group of L02 cell was treated with the same concentrations of SeMet as above for 48 h.The cell supernatant and lysates werecollected for the analysis of SELENOP and CPXI,respectively by WB.RESULTS The expressions of P-AKT-(Ser-473),P-AKT-(Thr-308),PI3K and mTOR in L02 cells under high-Se were decreased with the increase of SeMet concentration.The expressions of GPX1 and SELENOP were enhanced with the increase of SeMet.CONCLUSION The insulin signaling pathway,PI3K-AKT-mTOR,was damaged in L02 cell under high-Se stress.

关 键 词：硒 L02细胞 PI3K-AKT-mTOR 谷胱甘肽过氧化物酶1 硒蛋白P 

分 类 号：R151[医药卫生—营养与食品卫生学]