彭晶,郑锦坤,何蓉蓉,彭玉梅,李庆德
PENG Jing,ZHENG Jinkun,HE Rongrong,PENG Yumei,LI Qingde

• 1:粤北人民医院

摘要(Abstract)：

目的研究槲皮素对棕榈酸(PA)诱导的胰岛素抵抗HepG2细胞糖代谢的改善作用。方法用棕榈酸诱导的HepG2细胞建立胰岛素抵抗模型。用MTT法观察不同浓度槲皮素对HepG2细胞的影响,用葡萄糖氧化酶法检测葡萄糖消耗,用实时PCR(Real-time PCR)的方法检测AKT、GSK-3β、G6Pase、GCK的基因表达,用免疫印迹法(Western blotting)检测AKT、GSK-3β的磷酸化蛋白表达及G6Pase、GCK的蛋白表达。结果槲皮素能够增加棕榈酸诱导HepG2细胞中的葡萄糖消耗,明显增加棕榈酸诱导HepG2细胞中AKT的磷酸化水平,降低GSK-3β磷酸化水平,降低G6Pase基因和蛋白表达,增加GCK基因和蛋白表达。结论槲皮素能改善棕榈酸诱导的胰岛素抵抗HepG2细胞的糖代谢作用,与其激活AKT信号通路密切相关。
OBJECTIVE To investigate the improving effect of quercetin on glucose metabolism in PA-induced insulin resistant HepG2 cells. METHODS HepG2 cells were treated by PA to induce insulin resistance model. MTT assay was used to observe the influence of different concentration of quercetin on HepG2 cells. Glucose oxidation method was used to measure glucose consumption.Real-time PCR assay was used to detect the RNA expression levels of AKT,GSK-3β,G6 Pase and GCK. Western blotting assay was used to detect the phosphorylation levels of AKT and GSK-3β,and the protein levels of G6 Pase and GCK. RESULTS Quercetin effectively increased glucose consumption in PA-induced HepG2 cells. Quercetin markedly increased the phosphorylation levels of AKT,mRNA and protein levels of GCK,decreased the phosphorylation levels of GSK-3β,mRNA and protein levels of G6 Pase in PA-induced HepG2 cells.CONCLUSION The study confirmed that the improving effect of quercetin on glucose metabolism in PA-induced insulin resistant HepG2 cells is closely relevant to its role in activating AKT pathway.

关键词(KeyWords)： 槲皮素;胰岛素抵抗;AKT
quercetin;insulin resistance;AKT

Abstract:

Keywords:

基金项目(Foundation): 广东省医院药学研究基金(2016A02);; 广东省医学科学技术研究基金(A2016591)

作者(Author): 彭晶,郑锦坤,何蓉蓉,彭玉梅,李庆德
PENG Jing,ZHENG Jinkun,HE Rongrong,PENG Yumei,LI Qingde

参考文献(References)：

• [1]Zhang N,Du S M,Ma G S.Current lifestyle factors that increase risk of T2DM in China[J].Eur J Clin Nutr,2017,71(7):832-838.
• [2]Goodpaster B H,Sparks L M.Metabolic Flexibility in Health and Disease[J].Cell Metab,2017,25(5):1027-1036.
• [3]Kubota T,Kubota N,Kadowaki T.Imbalanced Insulin Actions in Obesity and Type 2 Diabetes:Key Mouse Models of Insulin Signaling Pathway[J].Cell Metab,2017,25(4):797-810.
• [4]虎松艳,王晓东,姜笑寒.灰树花菌丝体多糖对STZ致糖尿病小鼠血糖和血脂的影响[J].今日药学,2016,26(3):159-161.
• [5]向婷,何晓恩,潘卫松.番石榴叶及棠梨降血糖作用的实验研究[J].今日药学,2016,26(3):145-152.
• [6]Alam M M,Meerza D,Naseem I.Protective effect of quercetin on hyperglycemia,oxidative stress and DNA damage in alloxan induced type 2 diabetic mice[J].Life Sci,2014,109(1):8-14.
• [7]Xie X,Peng J,Huang K,et al.Polydatin ameliorates experimental diabetes-induced fibronectin through inhibiting the activation of NFkappa B signaling pathway in rat glomerular mesangial cells[J].Mol Cell Endocrinol,2012,362(1-2):183-193.
• [8]Hao J,Chen C,Huang K,et al.Polydatin improves glucose and lipid metabolism in experimental diabetes through activating the Akt signaling pathway[J].Eur J Pharmacol,2014,745:152-165.
• [9]Yin J,Hu R,Chen M,et al.Effects of berberine on glucose metabolism in vitro[J].Metabolism,2002,51(11):1439-1443.
• [10]张红,毕艳,葛智娟,等.促红细胞生成素通过叉头状转录因子O1-糖原合酶激酶3β信号改善棕榈酸诱导HepG2细胞糖代谢[J].中华糖尿病杂志,2016,8(3):157-161.
• [11]熊小侃,徐焱成,尚桂莲,等.过表达NR2E1对棕榈酸诱导的NIT1细胞凋亡及内质网应激的影响[J].中国糖尿病杂志,2016,24(8):733-737.
• [12]杨菊红,陈莉明,常宝成,等.SIRT1对高糖高脂培养胰岛微血管内皮细胞e NOS表达及活性的影响[J].中华内分泌外科杂志,2016,10(3):196-200.
• [13]Gonzalez E,Mc Graw T E.Insulin-modulated Akt subcellular localization determines Akt isoform-specific signaling[J].Proc Natl Acad Sci U S A,2009,106(17):7004-7009.
• [14]Manning B D,Toker A.AKT/PKB Signaling:Navigating the Network[J].Cell,2017,169(3):381-405.
• [15]Liu T Y,Shi C X,Gao R,et al.Irisin inhibits hepatic gluconeogenesis and increases glycogen synthesis via the PI3K/Akt pathway in type 2 diabetic mice and hepatocytes[J].Clin Sci,2015,129(10):839-850.
• [16]Japtok L,Schmitz E I,Fayyaz S,et al.Sphingosine 1-phosphate counteracts insulin signaling in pancreatic beta-cells via the sphingosine 1-phosphate receptor subtype 2[J].FASEB J,2015,29(8):3357-3369.
• [17]Lee J,Kim M S.The role of GSK3 in glucose homeostasis and the development of insulin resistance[J].Diabetes Res Clin Pract,2007,77(3):49-57.
• [18]Cohen P,Goedert M.GSK3 inhibitors:development and therapeutic potential[J].Nat Rev Drug Discov,2004,3(6):479-487.
• [19]Lin H V,Accili D.Hormonal regulation of hepatic glucose production in health and disease[J].Cell Metab,2011,14(1):9-19.
• [20]方达飞,王小琴,王长江,等.辛伐他汀联合依折麦布对2型糖尿病合并动脉粥样硬化大鼠糖脂及VEGF、IL-1β、CRP水平的影响[J].中国现代应用药学,2017,34(1):49-52.
• [21]常秀亭,李婕,谢曦,等.糖脉交泰胶囊对高糖高脂糖尿病大鼠糖脂代谢及肝脏葡萄糖激酶、低密度脂蛋白受体的影响[J].中国药理学通报,2013,29(2):234-236.