白子菜对HepG2细胞胰岛素抵抗作用的分子机制  

Molecular Mechanism of Gynura divaricata on Insulin Resistance in HepG2 Cells

作　　者：韦乃球[1,2] 郝二伟 冼寒梅[1] 覃文慧 覃骊兰[1] 李琦[1] 柳俊辉[1] 

机构地区：[1]广西中医药大学,南宁530200 [2]广西中药药效研究重点实验室,南宁530200

出　　处：《中国实验方剂学杂志》2018年第2期110-115,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广西自然科学基金青年基金项目(2014GXNSFBA118159);广西中药药效研究重点实验室系统性研究课题项目(14-B-03)

摘　　要：目的：观察白子菜对胰岛素抵抗人肝癌HepG2细胞的胰岛素受体（insulin receptor，InsR），葡萄糖转运蛋白4（glucosetransporter4，GLUT4）mRNA表达和蛋白激酶B（proteinkinaseB，PKB），糖原合成激酶-3／3（glycogensynthasekinase-3β，GSK一3／3）蛋白表达的影响，探讨其干预胰岛素抵抗的分子机制。方法：用胰岛素诱导HepG2细胞使其产生胰岛素抵抗，实验分为空白组，模型组，自子菜水提物（Gynura divaricata hot water extracts，GDE）高、低质量浓度（1．0，0．5g·L^-1）组，白子菜总黄酮（G．divaricataflavonoids，GDF）高、低质量浓度（0．2，0．1g·L^-1）组，白子菜总生物碱（G．divaricata alkaloid，GDA）高、低质量浓度（0．1，0．05g·L^-1）组，二甲双胍（1×10^-3mol·L^-1）组，葡萄糖氧化酶法检测HepG2细胞培养液上清液中葡萄糖的含量，实时荧光定量聚合酶反应（Real-timePCR）检测HepG2细胞InsR，GLUT4mRNA表达，蛋白免疫印迹法（Western blot）检测HepG2细胞PKB，GSK-3β的蛋白表达。结果：与模型组比较，1．0，0．5g·L。GDE组上清液葡萄糖含量极显著降低（P〈0．01），0．2g·L^-1GDF组上清液葡萄糖含量显著降低（P〈0．05）；GDE，GDF组InsR，GLUT4的mRNA表达显著增加（P〈0．05）；GDE组PKB的蛋白表达显著增加（P〈0．05），GSK-3β的蛋白表达显著降低（P〈0．05），GDF组PKB的蛋白表达极显著增加（P〈0．01），GSK-3β的蛋白表达极显著降低（P〈0．01）。结论：白子菜可改善胰岛素抵抗HepG2细胞对葡萄糖的摄取，其机制可能与上调胰岛素抵抗HepG2细胞InsR，GLUT4mRNA表达和PKB的蛋白表达及降低GSK-3β的蛋白表达相关。Objective： To observe the effect of Gynura divaricata on mRNA expressions of insulin receptor （InsR） and glucose transporter 4 （GLUT4） , and protein expressions of protein kinase B （PKB） and glycogen synthase kinase-3β （GSK-3β） on insulin resistance in HepG2 cells, in order to discuss the molecular mechanism of insulin resistance after intervention. Method： Insulin-induced HepG2 cells were used to produce insulin resistance. The experiment included blank control group, model group, high-dose and low-dose G. divaricata hot water extracts （ GDE, 1.0, 0.5 g·L-1 ） groups, high-dose and low-dose G. divaricata flavonoids （GI）F, 0.2, O. 1 g-L-1） groups, high-dose and low-dose G. divaricata alkaloid （ GI）A, 0.1, 0.05 g·L-1 ） groups. Glucose oxidase method was used to detect the content of glucose in supernatant of I-lepG2 cell culture medium; mRNA expressions of insulin receptor （InsR） and glucose transporter 4 （GLUT4） in HepG2 cells were detected by Real-time PCR; and Western blot was used to detect protein expressions of kinase B （PKB） and GSK-3fl in HepG2 cells. Result： Compared with model control group, the glucose content of supernatant was significantly decreased in 1.0, 0. 5 g.L-l GDE groups （P 〈 0. 01 ）, and the glucose content was significantly decreased in supernatant ofO. 2 g＇L-1GDF group （P 〈0. 05） ; the mRNA expressions of INSR and GLUT4 in GDE and GDF groups increased significantly （ P 〈 0.05 ） （P 〈 0.05） , and the protein expression of GSK-3fl protein expression of PKB was significantly increased ; the protein expression of PKB was significantly increased was significantly reduced in GDE group （ P 〈 0.05 ） , the （P 〈 0.01） and the protein expression of GSK-3fl decreased significantly in GDF group （ P 〈 0. 01 ）. Conclusion ： Gynura divaricata may improve insulin resistance HepG2 cells＇ uptake of glucose. Its mechanism may be related to the up-regulation of InsR, GLUT4 mRNA expression and PKB protein express

关 键 词：糖尿病 白子菜 HEPG2细胞 胰岛素 分子机制 

分 类 号：R285.5[医药卫生—中药学] R259[医药卫生—中医学]