摘要
目的探讨碧萝芷(PYC)对油酸(OA)诱导的AML12细胞胰岛素抵抗的作用及其相关分子机制。方法将细胞分为空白对照组、PYC50(50μg/mL)组、OA(200μmol/L)组、PYC10(10μg/mL)+OA(200μmol/L)组、PYC50(50μg/mL)+OA(200μmol/L)组。应用葡萄糖氧化酶法检测葡萄糖摄取量,糖原染色法检测糖原含量。实时PCR检测糖异生基因G6PC、PEPCK、PGC1αmRNA和糖酵解基因Eno1、PKLR、Gpi1 mRNA表达,Western blotting检测IRS/p-IRS、AKT/p-AKT、p-GSK3β的表达。结果与OA组比较,PYC10+OA组、PYC50+OA组培养基上清葡萄糖剩余含量明显减少(P<0.05),糖原含量明显增加(P<0.05)。PYC10+OA组、PYC50+OA组分别与OA组比较,糖异生基因G6PC、PEPCK、PGC1αmRNA表达无统计学差异(P>0.05),糖酵解基因Eno1、Gpi1 mRNA表达无统计学差异(P>0.05),但PKLR mRNA表达增加(P<0.05)。在胰岛素信号通路中,PYC10+OA组、PYC50+OA组分别与OA组比较,p-IRS表达无统计学差异(P>0.05),但p-AKT、p-GSK3β蛋白表达明显升高(P<0.05)。结论PYC通过激活AKT促进PKLR表达增加糖酵解,并且促进p-GSK3β表达增加糖原合成来调控AML12细胞胰岛素抵抗。
Objective To investigate the protective effect of pycnogenol(PYC)on oleic acid(OA)-induced insulin resistance in AML12 cells and its underlying mechanisms of action.Methods AML12 cells were divided into the blank control group,PYC50(50μg/mL)group,OA(200μmol/L)group,PYC10(10μg/mL)+OA(200μmol/L)group,and PYC50(50μg/mL)+OA(200μmol/L)group.Glucose uptake was examined using the glucose oxidase assay,and glycogen content was detected by PAS staining.The mRNA expression of the genes involved in gluconeogenesis(G6PC,PEPCK,and PGC1α)and glycolysis(Eno1,PKLR,and Gpi1)was detected by real time-PCR.The levels of proteins involved in the insulin signaling pathway(IRS/p-IRS,AKT/p-AKT,and p-GSK3β)were detected by Western blotting.Results The residual glucose content was significantly decreased(P<0.05)and the glycogen content was significantly increased in the PYC10+OA and PYC50+OA groups compared to the OA group(P<0.05).In comparison to the OA group,there was no significant difference in the expression of G6PC,PEPCK,PGC1α,Eno1,and Gpi1 mRNA(P>0.05),while PKLR mRNA expression was significantly increased(P<0.05)in the PYC10+OA and PYC50+OA groups.Analysis of insulin signaling pathway activation revealed that the expression of p-IRS was unaltered in the PYC10+OA and PYC50+OA groups(P>0.05),while the expression of p-AKT and p-GSK3βwas significantly increased(P<0.05)when compared to the OA group.Conclusion PYC increased the expression of PKLR and p-GSK3βby activating AKT,which further increased glycolysis and promoted glycogen synthesis,thus improving the OA-induced insulin resistance in AML12 cells.
作者
钟文霞
王艺颖
范斌
谷剑秋
ZHONG Wenxia;WANG Yiying;FAN Bin;GU Jianqiu(Department of Endocrinology and Metabolism,The First Hospital,China Medical University,Shenyang 110001,China;Clinical Nutrition Department,The Second Hospital of Wenzhou Medical University,Wenzhou 310009,China;Department of Neurology,Shengjing Hospital,China Medical University,Shenyang 110004,China)
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2020年第1期16-20,共5页
Journal of China Medical University
基金
国家自然科学基金（81100216）
关键词
碧萝芷
胰岛素抵抗
糖异生
糖酵解
pycnogenol
insulin resistance
gluconeogenesis
glycolysis
