基于IRS-1/PI3K/AKT/GLUT4通路探讨二苯乙烯类化合物改善胰岛素抵抗作用机制  

Mechanism of stilbene compounds on the improvement of insulin resistance by IRS-1/PI3K/AKT/GLUT4 pathway

作　　者：徐一方 李志鹏 曹世杰 康宁 XU Yifang;LI Zhipeng;CAO Shijie;KANG Ning(School of Integrative Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;State Key Laboratory of Component鄄based Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学中西医结合学院,天津301617 [2]天津中医药大学组分中药国家重点实验室,天津301617

出　　处：《天津中医药大学学报》2023年第1期95-102,共8页Journal of Tianjin University of Traditional Chinese Medicine

基　　金：国家自然科学基金重点项目(81430095)。

摘　　要：[目的]探讨3种中药(虎杖、桑白皮、何首乌)来源的二苯乙烯类化合物改善胰岛素抵抗(IR)的作用机制。[方法]利用肿瘤坏死因子-α(TNF-α)作用于3T3-L1脂肪细胞96 h建立IR细胞模型;采用噻唑蓝(MTT)法检测二苯乙烯类化合物白藜芦醇(RES)、虎杖苷(PD)、桑皮苷A(Mul A)、2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡糖糖苷(THSG)对脂肪细胞生存率的影响;葡萄糖氧化酶法检测细胞培养基上清液中葡萄糖含量;采用蛋白免疫印迹(Western Blot)法检测胰岛素受体底物-1(IRS-1)、磷酸化IRS-1(p-IRS-1)、磷脂酰肌醇-3激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、葡萄糖转运体4(GLUT4)的蛋白表达水平;采用实时定量荧光聚合酶链反应(RT-PCR)法检测GLUT4 mRNA表达水平。[结果]与空白对照组相比,模型组的葡萄糖消耗量显著下降,模型组的p-IRS-1(Ser307)蛋白表达显著增加,p-PI3K、p-AKT(Ser473)、GLUT4蛋白表达显著下降,模型组GLUT4 mRNA表达显著降低;与模型组相比,RES、Mul A、THSG组的葡萄糖消耗量显著增加,PD组的葡萄糖消耗量无明显变化,Mul A、THSG组均能显著逆转p-IRS-1(Ser307)、p-PI3K、p-AKT(Ser473)、GLUT4的蛋白表达,RES仅能够逆转p-IRS-1(Ser307)、GLUT4的蛋白表达;与模型组相比,THSG组GLUT4 mRNA表达显著增加。[结论]Mul A、THSG能够通过IRS-1/PI3K/AKT/GLUT4信号通路改善脂肪细胞IR,而RES则通过激活IRS-1和GLUT4,但不通过PI3K/AKT途径发挥改善脂肪细胞IR的作用。PD无降糖活性。[Objective]The aim of this study is to investigate the mechanism of three traditional Chinese medicines(Polygoni Cuspidati Rhizoma et Radix,Mori Cortex,Polygoni Multiflori Radix)derived stilbene compounds in improving insulin resistance.[Methods]The insulin resistance model was established by using tumor necrosis factorα(TNF-α)for 96 h in 3T3-L1 adipocytes.Thiazolyl blue tetrazolium bromide(MTT)method was used to detect the survival rates of adipocytes with stilbene compounds,such as resveratrol(RES),polydatin(PD),mulberroside A(Mul A)and 2,3,5,4’-tetrahydroxyl diphenylethylene-2-O-glucoside(THSG).Glucose content in supernatant of cell culture medium was detected by glucose kit.Protein expression levels of insulin receptor substrate-1(IRS-1),p-IRS-1,phosphatidylinositol3-kinase(PI3K),p-PI3K,protein kinase B(AKT),p-AKT,glucose transporter 4(GLUT4)were detected by Western Blot,and the mRNA expression of GLUT4 was detected by RT-PCR.[Results]Compared with those in the normal group,the glucose consumption rate of the model group was significantly down-regulated,meanwhile the protein expression of p-IRS-1(Ser307)was significantly increased,the expressions of p-PI3K and pAKT(Ser473)and GLUT4 were down-decreased,and the mRNA expression of GLUT4 was significantly decreased.Compared with the model group,RES,Mul A,and THSG could significantly increase the glucose consumption,while PD has no such effect.Moreover,the protein expressions of p-IRS-1(Ser307)and p-PI3K and p-AKT(Ser473)and GLUT4 were significantly reversed by Mul A and THSG.However,RES only significantly reverse the protein expressions of p-IRS-1(Ser307)and GLUT4.THSG could significantly increase the mRNA expression of GLUT4.[Conclusion]Mul A and THSG could improve the insulin resistance via IRS-1/PI3K/AKT/GLUT4 signaling pathway in the adipocytes.RES could improve the insulin resistance by activating IRS-1 and GLUT4 but not through the PI3K/AKT pathway in the adipocytes,and PD has no hypoglycemic activity.

关 键 词：二苯乙烯 脂肪细胞 胰岛素抵抗 IRS-1/PI3K/AKT/GLUT4 

分 类 号：R285.5[医药卫生—中药学]