α-硫辛酸对胰岛素抵抗HepG2细胞糖代谢紊乱的改善作用  

Improvement Effects ofα-lipoic Acid on Glucose Metabolism Disorder of Insulin Resistant HepG2 Cells

作　　者：安仙蓉 管俊欢 杨秀丽 吴先进 梁冰 AN Xianrong;GUAN Junhuan;YANG Xiuli;WU Xianjin;LIANG Bing(Dept.of Pharmacology,School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Environmental Pollution and Disease Monitoring,Ministry of Education,Guiyang 550025,China)

机构地区：[1]贵州医科大学基础医学院药理学教研室,贵阳550025 [2]环境污染与疾病监控教育部重点实验室,贵阳550025

出　　处：《中国药房》2021年第14期1703-1708,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81760575)。

摘　　要：目的:研究α-硫辛酸对胰岛素抵抗HepG2细胞糖代谢紊乱的改善作用。方法:采用MTT法测定25~1000μmol/Lα-硫辛酸对人肝癌细胞HepG2存活率的影响,以确定α-硫辛酸给药浓度。实验设阴性对照组、胰岛素抵抗组(1×10-7 mol/L胰岛素)、联合抵抗组(30μmol/L亚砷酸钠+1×10-8 mol/L胰岛素)和α-硫辛酸低、中、高浓度组。按分组加入α-硫辛酸作用HepG2细胞12 h后,再分别给予相应浓度的亚砷酸钠或/和胰岛素继续培养24 h。采用葡萄糖氧化酶法检测细胞葡萄消耗量,比色法检测细胞己糖激酶、丙酮酸激酶活性,蒽酮法检测细胞糖原含量,Westernblot法检测细胞中葡萄糖转运体4(GLUT4)、磷酸化糖原合成激酶3β(p-GSK3β)、GSK3β蛋白表达水平和磷酸化蛋白激酶B(p-Akt)/Akt、p-GSK3β/GSK3β比值。结果:25、50、100μmol/L的α-硫辛酸对细胞存活率无明显影响(P>0.05),且细胞存活率均大于96%,故将其作为后续研究的低、中、高浓度。与阴性对照组比较,胰岛素抵抗组、联合抵抗组细胞葡萄糖消耗量、己糖激酶和丙酮酸激酶活性、糖原含量、GLUT4和p-GSK3β蛋白表达水平、p-Akt/Akt和p-GSK3β/GSK3β比值均显著降低,GSK3β蛋白表达水平均显著升高(P<0.05)。与联合抵抗组比较,α-硫辛酸各浓度组细胞葡萄糖消耗量(α-硫辛酸低浓度组除外)、己糖激酶(α-硫辛酸低、中浓度组除外)和丙酮酸激酶(α-硫辛酸低、中浓度组除外)活性、糖原含量、GLUT4(α-硫辛酸低浓度组除外)、p-GSK3β蛋白表达水平和p-Akt/Akt(α-硫辛酸低、中浓度组除外)、p-GSK3β/GSK3β(α-硫辛酸低浓度组除外)比值均显著升高,GSK3β(α-硫辛酸低、中浓度组除外)蛋白表达水平均显著降低(P<0.05),且α-硫辛酸高浓度组细胞糖原含量、GLUT4蛋白表达水平、p-GSK3β/GSK3β比值和α-硫辛酸中、高浓度组细胞p-GSK3β蛋白表达水平的改善效果更明显(P<0.05)。结论:α-硫辛酸对胰岛素抵抗HeOBJECTIVE:To study the improvement effects ofα-lipoic acid on glucose metabolism disorder of insulin resistant HepG2 cells.METHODS:The effects of 25-1000μmol/Lα-lipoic acid on survival rate of human hepatoma cell HepG2 were determined by MTT assay so as to determine the concentration ofα-lipoic acid.Negative control group,insulin resistance group(1×10-7 mol/L insulin),combination resistance group(30μmol/L sodium arsenite+1×10-8 mol/L insulin),α-lipoic acid low concentration,medium-concentration and high-concentration groups were set up.HepG2 cells were treated withα-lipoic acid for 12 h and then cultured with corresponding concentration of sodium arsenite or/and insulin for 24 h.The glucose oxidase method was used to detect the glucose consumption,colorimetric method was used to detect hexokinase activity and pyruvate kinase activity,and anthrone method was used to detect glycogen content.Western blot assay was used to detect the protein expression of GLUT4,p-GSK3βand GSK3βas well as the ratio of p-Akt/Akt and p-GSK3β/GSK3β.RESULTS:25,50,100μmol/Lα-lipoic acid had no significant effect on the survival rates of HepG2 cells(P>0.05),and survival rates of HepG2 cells were higher than 96%,so they were used as the low,medium and high concentration for follow-up study.Compared with negative control group,glucose consumption,the activities of hexokinase and pyruvate kinase,glycogen content,protein expression of GLUT4 and p-GSK3β,the ratio of p-Akt/Akt and p-GSK3β/GSK3βwere decreased significantly in insulin resistance group and combined resistance group,while the protein expression of GSK3βwas increased significantly(P<0.05).Compared with combination resistance group,the glucose consumption(except forα-lipoic acid low-concentration group),the activities of hexokinase(except forα-lipoic acid low-concentration and medium-concentration groups)and pyruvate kinase(except forα-lipoic acid low-concentration and medium-concentration groups),glycogen contents,protein expression of GLUT4(except forα-lipoic a

关 键 词：Α-硫辛酸 人肝癌细胞HEPG2 亚砷酸钠 胰岛素抵抗 糖代谢紊乱 磷脂酰肌醇3-激酶/蛋白激酶B 

分 类 号：R965[医药卫生—药理学] Q493.4[医药卫生—药学]