葛根总黄酮对胰岛素抵抗模型HepG2细胞糖代谢的改善作用及机制研究  

Effects of Crude Flavones of Pueraria lobata on the Improvement of Glycometabolism in Insulin-resistant HepG2 Cells and Underlying Mechanism

作　　者：王茹 袁雅倩 曾敏[3] 陈慧冬 Wang Ru;Yuan Yaqian;Zeng Min(Department of Geratology,China Resources and WISCO General Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430080,China;Medical College,Wuhan University of Science and Technology,Wuhan 430062,China;Department of Endocrinology,Puren Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430081,China)

机构地区：[1]武汉科技大学附属华润武钢总医院老年病科,武汉430080 [2]武汉科技大学医学院,武汉430062 [3]武汉科技大学附属普仁医院内分泌科,武汉430081 [4]武汉市金银潭医院呼吸介入科,武汉430023

出　　处：《华中科技大学学报（医学版）》2023年第1期43-47,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：武汉市卫生计生委临床医学科研基金资助项目(No.WX18D11,No.WZ17B09);湖北省卫生计生委联合基金资助项目(No.WJ2018H0114)。

摘　　要：目的研究葛根总黄酮对胰岛素抵抗(IR)HepG2细胞模型糖代谢的影响,并探讨其作用机制。方法采用含胰岛素0.1μmol/L的培养液诱导HepG2细胞IR模型,使用10、20和50μmol/L葛根总黄酮干预细胞,筛选出最佳浓度。Western blot检测各组细胞内葡萄糖转运蛋白4(GLUT4)、泛素E3连接酶(c-Cbl)、磷脂酰肌醇三羟基激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)和磷酸化AKT(p-AKT)蛋白表达水平;比色法测定细胞内葡萄糖、乳酸及ATP含量;蒽酮法测定细胞内糖原含量。结果成功构建HepG2细胞IR模型。与对照组比较,模型组细胞内GLUT4、c-Cbl、p-PI3K和p-AKT蛋白表达水平显著降低(均P<0.01),葡萄糖、乳酸、ATP及糖原含量也显著降低(均P<0.01)。50μmol/L葛根总黄酮干预IR HepG2细胞模型24 h后,细胞内GLUT4、c-Cbl、p-PI3K和p-AKT蛋白表达升高(均P<0.01),且细胞内葡萄糖、乳酸、ATP及糖原含量升高(均P<0.01)。结论葛根总黄酮可提高IR模型HepG2细胞对胰岛素的敏感性,促进葡萄糖摄取及细胞内的糖代谢过程,其作用机制可能与上调c-Cbl/PI3K/AKT信号通路相关。Objective To investigate the effect and mechanism of crude flavones of Pueraria lobata on the glycometabolism of HepG2 cells in an insulin resistance(IR)model.Methods The IR model of HepG2 cell was induced by medium containing 0.1μmol/L insulin.The best concentration of crude flavones of Pueraria lobata was screened by treating the cells with 10,20 and 50μmol/L insulin.The protein expression of glucose transporter 4(GLUT4),casitas B-lineage lymphoma(c-Cbl),phosphoinositide 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),and phosphorylated AKT(p-AKT)were detected by Western blotting.Cellular glucose content,lactate production,and ATP levels were measured by colorimetric assays,and the content of glycogen was determined by the anthrone method.Results The IR model was successfully established in HepG2 cells.Compared with the control group,the expression levels of GLUT4,c-Cbl,p-PI3K,and p-AKT in the model group were significantly decreased(all P<0.01).Glucose,lactate,ATP,and glycogen content were also significantly reduced(all P<0.01).Twenty-four hours after IR HepG2 cells were treated with crude flavones of Pueraria lobata,the expression levels of GLUT4,c-Cbl,p-PI3K and p-AKT were increased(all P<0.01),glucose,lactate,ATP and glycogen levels were increased(all P<0.01).Conclusion Crude flavones of Pueraria lobata can enhance the sensitivity of IR HepG2 cells to insulin and promote glucose uptake and intracellular glucose metabolism.Its mechanism may be related to upregulation of the c-Cbl/PI3K/AKT signaling pathway.

关 键 词：葛根总黄酮 胰岛素抵抗 糖代谢 葡萄糖转运蛋白4 c-Cbl/PI3K/AKT信号通路 

分 类 号：R587.1[医药卫生—内分泌]