LI Xiu-li1, HE Song-min2, ZHU Ying2, et al. Establishment and Identify of HepG2 Cells Model of Insulin Resistance[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(5): 203-207.
LI Xiu-li1, HE Song-min2, ZHU Ying2, et al. Establishment and Identify of HepG2 Cells Model of Insulin Resistance[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(5): 203-207. DOI: 10.13422/j.cnki.syfjx.2013.05.101.
Establishment and Identify of HepG2 Cells Model of Insulin Resistance
酶联免疫法检测葡萄糖-6-磷酸酶(G-6-Pase)的表达。结果:细胞产生胰岛素抵抗的最佳作用时间为36 h
最佳胰岛素浓度为1×10-8mol.L-1。建立模型后鉴定
模型细胞与正常细胞相比脂滴明显增多;IRS-2 mRNA的表达较正常细胞明显降低(P<0.05)
G-6-Pase的表达较正常细胞明显升高(P<0.05)。结论:采用胰岛素体外诱导的方法
在胰岛素浓度为1×10-8mol.L-1
作用36 h后
可建立稳定的HepG2细胞胰岛素抵抗模型。
Abstract
Objective:To establish the HepG2 cells model of insulin resistance by the method of insulin-induced in vitro
and provide a reliable model of insulin resistance in vitro for the study of the insulin resistance and the type 2 diabetes.Method: RPMI 1640 medium with different insulin concentrations was used to the HepG2 cells
the medium glucose content was detected at different time points by the method of glucose-hexokinase and the HepG2 cell viability of different insulin concentrations was detected by the method of MTT
and to determine the optimal concentration and time of the insulin resistance;then the changes of cell morphology was observed by the oil red O staining;the expression of insulin receptor substance(IRS)-2 mRNA was detected by the method of real-time PCR and the expression of G-6-Pase was detected by the method of ELISA.Result: The best time of insulin resistance was 36 h
the best concentration of insulin resistance was 1×10-8 mol·L-1.After the establishment of the model
the lipid droplets of model cells was significantly increased compared with normal cells;the IRS-2 mRNA expression was significantly reduced compared with normal cells(P<0.05);the G-6-Pase expression was significantly increased compared with normal cells(P<0.05).Conclusion: By the method of insulin-induced in vitro
a stable HepG2 insulin resistance model can be established when the 1×10-8mol·L-1 concentration of insulin act on the HepG2 cell for 36 h.
Establishment and Identify of HepG2 Cells Model of Insulin Resistance
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Related Author
ZHENG Guang-juan
Chen Ting
HUANG Xiao-qian
FENG Bing
ZHU Ying
HE Song-min
LI Xiu-li
Meng WANG
Related Institution
State Key Laboratory of Component-based Chinese Medicine, Institute of Traditional Chinese Medicine (TCM),Tianjin University of TCM
The First Affiliated Hospital of Chongqing Medical University
College of Pharmacy and Chemistry, Key Laboratory of Universities in Yunnan Province on Development of Genuine Medicinal Materials Resources in Western Yunnan, Dali University