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α-硫辛酸对胰岛素抵抗HepG2细胞糖代谢紊乱的改善作用
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| 篇名： | α-硫辛酸对胰岛素抵抗HepG2细胞糖代谢紊乱的改善作用 |
| TITLE： | Improvement Effects of α-lipoic Acid on Glucose Metabolism Disorder of Insulin Resistant HepG 2 Cells |
| 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（1×10－7mol/L胰岛素）、联合抵抗组（30µmol/L亚砷酸钠+1×10－8mol/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12h后，再分别给予相应浓度的亚砷酸钠或/和胰岛素继续培养24h。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HepG2细胞的糖代谢紊乱具有一定的改善作用，其机制可能与增加葡萄糖消耗，增强糖代谢相关酶活性，提高糖原含量，上调Akt、GSK3β蛋白的磷酸化水平和GLUT4、p-GSK3β蛋白的表达水平，下调GSK3β蛋白的表达水平有关。 |
| ABSTRACT： | OBJECTIVE：To study the i mprovement effects of α-lipoic acid on glucose metabolism disorder of insulin resistant HepG2 cells. METHODS ：The effects of 25-1 000 µmol/L α-lipoic acid on survival rate of human hepatoma cell HepG2 were determined by MTT assay so as to determine the concentration of α-lipoic acid. Negative control group ，insulin resistance group （1× 10－7 mol/L insulin ），combination resistance group （30 µmol/L sodium arsenite+ 1×10－8 mol/L insulin ），α-lipoic acid low- concentration，medium-concentration and high-concentration groups were set up. HepG 2 cells were treated with α-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 GLUT 4， p-GSK3β and GSK3β as well as the ratio of p-Akt/Akt and p-GSK3β/GSK3β. RESULTS：25，50，100 µmol/L α-lipoic acid had no significant effect on the survival rates of HepG 2 cells（P＞0.05），and survival rates of H epG2 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 GLUT 4 and p-GSK 3β，the ratio of p-Akt/Akt and p-GSK 3β/GSK3β were decreased significantly in insulin resistance group and combined resistance group， while the protein expression of GSK 3β was increased significantly（P＜0.05）. Compared with combination resistance group ，the glucose consumption （except for α-lipoic acid low- concentration group ），the activities of h exokinase（except for α-lipoic acid low-concentration and medium-concentration groups ） andpyruvate kinase （except for α-lipoic acid low-concentration com and medium-concentration groups ）， glycogen contents ， protein expression of GLUT 4 （except for α-lipoic acid mail：bliang163@163.com low-concentration group ）and p-GSK3β，the ratio of p-Akt/ Akt（except for α-lipoic acid low-concentration and medium-concentration groups ）and p-GSK 3β/GSK3β（except for α-lipoic acid low-concentration groups ）were increased significantly in α-lipoic acid groups ，while protein expression of GSK 3β（except for α-lipoic acid low-concentration and medium-concentration groups ） was decreased significantly （P＜0.05）；glycogen content ， protein expression of GLUT 4 and the ratio of p-GSK 3β/GSK3β in α-lipoic acid high-concentration group as well as the protein expression of p-GSK 3β in α-lipoic acid medium-concentration and high-concentration groups were improved significantly （P＜0.05）. CONCLUSIONS：α-lipoic acid can improve the disorder of glucose metabolism in insulin resistant HepG 2 cells，the mechanism of which may be associated with the increase of glucose consumption ，the activities of glucose metabolism related enzymes and glycogen content ，and expression up-regulation of the phosphorylation levels of Akt and GSK 3β protein，the expression of GLUT 4 and p-GSK 3β proteins，down-regulation of the expression of GSK 3β protein. |
| 期刊： | 2021年第32卷第14期 |
| 作者： | 安仙蓉，管俊欢，杨秀丽，吴先进，梁冰 |
| AUTHORS： | AN Xianrong ，GUAN Junhuan ，YANG Xiuli，WU Xianjin ，LIANG Bing |
| 关键字： | 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 |
| KEYWORDS： | α-lipoic acid ；Human hepatoma cell HepG 2；Sodium arsenite ；Insulin resistance ；Glucose metabolism disorder ； |
| 阅读数： | 478 次 |
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