摘要
目的:探讨葛根芩连汤含药血清改善HepG2细胞胰岛素抵抗的机制。方法:建立胰岛素抵抗HepG2细胞模型,用葛根芩连汤含药血清、吡格列酮干预,并设空白对照组和模型组。比色法检测细胞糖原合成能力,实时定量PCR检测miR-146b的表达,Western blot和实时定量PCR检测SIRT1/FoxO1通路中相关蛋白及mRNA的表达。结果:葛根芩连汤含药血清能逆转胰岛素抵抗HepG2细胞的糖原合成能力下降(P<0.01)。与对照组比较,模型组miR-146b表达显著升高(P<0.01),葛根芩连汤含药血清能显著抑制miR-146b的表达(P<0.01)。Western blot显示,与空白对照组比较,模型组SIRT1、PPARγ蛋白表达显著降低,acetyl-FoxO1蛋白表达显著升高(P<0.05或P<0.01);与模型组比较,葛根芩连汤含药血清组SIRT1、PPARγ蛋白表达显著升高,acetyl-FoxO1蛋白表达显著降低(P<0.05或P<0.01)。过表达的miR-146b能抑制SIRT1蛋白的表达,沉默miR-146b能增强SIRT1蛋白的表达(P<0.05或P<0.01)。结论:葛根芩连汤含药血清能改善棕榈酸诱导的HepG2胰岛素抵抗模型,其机制与调控miR-146b/SIRT1信号通路有关。
Objective:To explore the mechanism of drug serum from Gegen qinlian decoction in improving insulin resistant in HepG2 cells.Methods:The insulin resistant HepG2 cell model was established.Drug serum from Gegen qinlian decoction and pioglitazone were used to intervene.Normal control group and model group were set up.The ability of glycogen synthesis was detected by colorimetry,the expression of miR-146 b was detected by real-time PCR,and the expression of related proteins and mRNAs in SIRT1/FoxO1 pathway was detected by Western blot and real-time PCR.Results:Drug serum from Gegen qinlian decoction could reverse the decline of glycogen synthesis in insulin resistant HepG2 cells(P<0.01).The expression of miR-146 b in model group was significantly increased compared with control group(P<0.01),and the expression of miR-146 b was significantly reduced by adding drug serum from Gegen qinlian decoction(P<0.01).Western blot showed that compared with normal control group,the expressions of SIRT1 and PPARγin model group decreased significantly,and the expression of acetyl-FoxO1 increased significantly(P<0.05 or P<0.01).Compared with model group,the expressions of SIRT1 and PPARγincreased significantly and the expression of acetyl-FoxO1 protein decreased(P<0.05 or P<0.01)in Gegen qinlian decoction drug serum group.Over-expressed miR-146 b could inhibit the expression of SIRT1 protein.Silencing miR-146 b could enhance the expression of SIRT1(P<0.05 or P<0.01).Conclusion:The drug serum from Gegen qinlian decoction can improve the insulin resistant HepG2 cells induced by palmitic acid.Its mechanism is related to the regulation of miR-146 b/SIRT1 signaling pathway.
作者
隋淼
蒋小飞
范尧夫
陈国芳
刘超
SUI Miao;JIANG Xiao-fei;FAN Yao-fu;CHEN Guo-fang;LIU Chao(Nanjing University of Traditional Chinese Medicine,Nanjing 210028,China;Xuzhou Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Xuzhou 221009,China;Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Traditional Chinese Medicine,Nanjing 210028,China)
出处
《中药材》
CAS
北大核心
2019年第3期630-635,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金青年基金(81603585)
