光甘草定通过调节ERK/IRS-1和PI3K/Akt信号通路改善HepG2细胞的胰岛素抵抗  

Glabridin ameliorates insulin resistance by regulating ERK/IRS-1 and PI3K/Akt signaling pathways in HepG2 cells

作　　者：李德锋 樊金玲[1] 杜琳[1] 任国艳[1] LI De-feng;FAN Jin-ling;DU Lin;REN Guo-yan(College of Food and Bioengineering,Henan University of Science and Technology,Luoyang 471023,China)

机构地区：[1]河南科技大学食品与生物工程学院,河南洛阳471023

出　　处：《中草药》2022年第24期7751-7762,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(31571800)。

摘　　要：目的 探索光甘草定改善肝细胞胰岛素抵抗(insulin resistance,IR)的作用和机制。方法 通过高胰岛素诱导人肝癌HepG2细胞建立IR模型,采用葡萄糖氧化酶法检测细胞的葡萄糖消耗量及生成;荧光标记法检测葡萄糖摄取量;蒽酮法检测糖原含量;ELISA检测葡萄糖代谢关键酶的活性;Western blotting检测磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol3-kinase/protein kinase B,PI3K/Akt)、细胞外调节蛋白激酶/胰岛素受体底物-1(extracellular regulated protein kinase/insulin receptor substrate-1,ERK/IRS-1)信号通路相关蛋白以及葡萄糖转运蛋白4(glucose transporter 4,GLUT4)的表达。采用分子对接技术研究光甘草定和ERK分子间的相互作用。结果 光甘草定显著增加IR-HepG2细胞的葡萄糖消耗和摄取(P<0.05);通过显著提高糖原合成酶(glycogen synthase,GS)、葡萄糖激酶(glucokinase,GCK)和丙酮酸激酶(pyruvate kinase,PK)活性(P<0.05、0.01),促进IR-HepG2细胞的糖原合成和糖酵解;通过显著减弱磷酸烯醇丙酮酸羧激酶(phosphoenolpyruvate carboxykinase,PEPCK)和葡萄糖-6-磷酸酶(glucose-6-phosphatase,G6Pase)的活性(P<0.05),抑制IR-HepG2细胞的糖异生。IR-HepG2细胞经光甘草定处理后,Akt、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)和叉头框蛋白O1(forkhead boxing protein O1,FOXO1)的磷酸化水平得到显著恢复(P<0.01),而这种作用被PI3K的抑制剂LY294002所逆转(P<0.01)。同时,光甘草定显著促进GLUT4向质膜的易位(P<0.01)。光甘草定显著降低IRHep G2细胞的ERK和IRS的磷酸化水平(P<0.01),还可作为ERK的I1/2型抑制剂。结论 光甘草定通过抑制ERK/IRS-1通路,激活PI3K/Akt信号通路,修复IR-HepG2细胞的糖代谢紊乱,缓解IR症状。Objective To explore the effect and mechanism of glabridin on ameliorating insulin resistance(IR) in hepatocytes.Methods IR model was established by high insulin-induced HepG2 cells.The cells were evaluated for glucose consumption and production by glucose oxidase assay;The glucose consumption and production of cells were detected by glucose oxidase method;Glucose uptake was detected by fluorescence method;The content of glycogen was detected by anthrone method;The activities of key enzymes in glucose metabolism was detected by ELISA;Western blotting was used to detect phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt),extracellular regulated protein kinase/insulin receptor substrate-1(ERK/IRS-1) signaling pathway related protein and glucose transporter 4(GLUT4) expressions.Molecular docking technique was used to study the interaction between glabridin and ERK molecules.Results Glabridin significantly increased the glucose consumption and uptake of IR-HepG2 cells(P < 0.05);Glycogen synthesis and glycolysis of IR-HepG2 cells were promoted by significantly increasing the activities of glycogen synthase(GS),glucokinase(GCK) and pyruvate kinase(PK)(P < 0.05,0.01);The activities of phosphoenolpyruvate carboxykinase(PEPCK)and glucose-6-phosphatase(G6Pase) were significantly decreased(P < 0.05),and gluconeogenesis of IR-HepG2 cells was inhibited.After IR-HepG2 cells were treated with glabridin,phosphorylation levels of Akt,glycogen synthase kinase-3β(GSK-3β) and forkhead boxing protein O1(FOXO1) were significantly restored(P < 0.01),and this effect was reversed by PI3K inhibitor LY294002(P <0.01).Meanwhile,glabridin significantly promoted the translocation of GLUT4 to plasma membrane(P < 0.01).Glabridin significantly reduced the phosphorylation levels of ERK and IRS in IR-HepG2 cells(P < 0.01),and could be used as I1/2 inhibitor of ERK.Conclusion Glabridin can repair the sugar metabolism disorder of IR-HepG2 cells and relieve IR symptoms by inhibiting ERK/IRS-1pathway and activating PI3K/Akt signaling pathway.

关 键 词：胰岛素抵抗 ERK/IRS-1 PI3K/Akt 光甘草定 HEPG2细胞 糖代谢 葡萄糖摄取 

分 类 号：R285.5[医药卫生—中药学]