枸杞多糖对HepG2细胞胰岛素抵抗的改善作用及机制研究  

Ameliorative effects and the mechanism of lyceum barbarum polysaccharide on insulin resistance of HepG2 cell

作　　者：许拓 凌宏艳[1] 龙佳 何剑琴[1] 杨丝丝[1] 朱责梅 颜灿群 奉水东[3] 

机构地区：[1]南华大学生理学教研室,湖南衡阳421001 [2]南华大学附属第二医院内分泌科,湖南衡阳421001 [3]南华大学社会医学与卫生事业管理学教研室,湖南衡阳421001

出　　处：《中国应用生理学杂志》2017年第6期568-571,共4页Chinese Journal of Applied Physiology

基　　金：教育部留学归国基金(教外司留[2014]1685号);湖南教育厅(16C1411);衡阳市科技局(2016KJ64)

摘　　要：目的:观察不同浓度的枸杞多糖(LBP)对Hep G2细胞胰岛素抵抗的影响并探讨其机制。方法:采用高糖高胰岛素处理Hep G2细胞24 h建立胰岛素抵抗细胞模型后,用台盼蓝检测活力大于95%的HepG2细胞,以104/孔密度接种于96孔板内,细胞贴壁后以30μg/ml、100μg/ml、300μg/ml的LBP培养48 h,200μl/well,各组均设4个复孔。检测不同浓度的LBP对Hep G2细胞活性及胰岛素抵抗的影响;细胞内丙二醛(MDA)含量和超氧化物歧化酶(SOD)的活性;各组细胞胰岛素信号转导通路中相关蛋白(IRS-2、PI3-K、Akt、GLUT2)的表达。结果:MTT显示:与正常对照组相比,IR模型组MDA含量显著升高,SOD活力明显降低,同时IRS-2、PI-3K、Akt、GLUT2蛋白表达水平明显下降;与IR模型组相比,中、高浓度LBP组MDA的含量明显降低,SOD的活力显著升高,且IRS-2、PI-3K、Akt、GLUT2蛋白表达水平明显升高;在相同的时间内,随着LBP浓度的增加,OD值逐渐降低;在同一浓度干预下,随着时间的延长,OD值也逐渐降低;葡萄糖消耗实验表明中、高浓度的LBP可显著提高胰岛素抵抗Hep G2细胞的葡萄糖消耗量,而低浓度LBP对Hep G2细胞葡萄糖消耗量无明显影响。结论:中、高浓度枸杞多糖能改善Hep G2细胞胰岛素抵抗,其作用机制可能与降低细胞氧化应激水平及提高胰岛素信号传导通路相关蛋白表达有关。Objective： To observe the effects of lyceum barbarum polysaeeharide （LBP） on insulin resistance of HepG2 cells and investigate its possible mechanism. Methods： IR-HepG2 cell model was induced with high glucose and high insulin in combination for 24 hours, with 104/vaccinatlon in the 96 - well plates, hole density after adherent cells （30 μg/ml, 100 μg/m1,300 μg./ml） LBP cultivate 48 h, 200 μl/hole, each all had four holes. The effects of LBP at difterent concentralions on HepG2 cell activity and insulin resistance were tested, Intraeellular malondialdehyde （MDA） content and supernxide dismutase （SOD） activity were detected. The expressions of related proteins in insulin signal transduction pathways such as insulin receptor substrate-2（ IRS-2）, phosphatidylinnsitol-3-kinase（Pl3-K）, protein kinase B（Akt） and glucose transport-2（GLUT2） were determined. Results： Compared with normal control group, the content of MDA was increased significantly and the aetivity of SOD and the expression levels of IRS-2, PI-3K, Akt and GLUT2 were decreased significantly in the IR model group. Compared with IR model group, meditum and high concentrations of LBP decreased the content of MDA and inereased the activity of SOD and the expression levels of IRS-2, PI-3K, Akt and GLUT2 in insulin-resistant HepG2 cells. MTT showed that at the same time, the OD value gradually decreased with the increase of LBP＇ s concentration; under the same concentration of LBP, the OD value also gradually decreased with the extension of time, which indicated that LBP inhibited the proliferation of HepG2 cells with time and coneentralion-dependent manner. Glucose consumption experiment indicated that medium and high concentration of LBP eouht increase the glueose consumption of insulin-resistant HepG2 cells significantly, hut low concentration of LBP had no significant impacted on glucose consumption of irLsulin-resistant HepC2 cells. Conclu- sion： Medium and high coneentration of I,BP ean improve insulin resistan

关 键 词：枸杞多糖 HEPG2细胞 胰岛素抵抗 氧化应激 胰岛素信号通路 

分 类 号：Q0[生物学] R742[医药卫生—神经病学与精神病学]