论文部分内容阅读
抵抗素是2001年Steppan等发现的一种与胰岛素抵抗有密切联系的细胞因子.本研究探讨了成纤维细胞生长因子21(FGF-21)在抵抗素过表达导致胰岛素抵抗的肝细胞中的糖代谢调节作用.构建人抵抗素真核表达载体,转染HepG2细胞,利用流式细胞仪筛选出过表达抵抗素的HepG2模型细胞,分别用不同浓度的胰岛素和FGF-21刺激细胞,用GOD-POD法检测细胞的葡萄糖摄取情况,利用实时荧光定量PCR方法检测抵抗素转染后及FGF-21处理后细胞GLUT1、PPAR-γmRNA表达的变化.PCR鉴定结果表明过表达抵抗素的HepG2模型细胞构建成功.GOD-POD法检测结果证明,模型细胞对胰岛素敏感性降低,但FGF-21仍能有效调节模型细胞的葡萄糖摄取,且呈现剂量依赖关系.实时荧光定量PCR方法检测发现,抵抗素转染后HepG2细胞GLUT1 mRNA表达增加,经FGF-21刺激后模型细胞与对照细胞相比GLUT1 mRNA的表达仍有上升趋势,PPAR-γ的变化不显著.上述结果表明,抵抗素过表达的肝细胞,对胰岛素敏感性降低,产生胰岛素抵抗,但FGF-21仍能有效调节其葡萄糖代谢.
Resistin is a cytokine closely related to insulin resistance discovered by Steppan et al in 2001. This study investigated the effect of fibroblast growth factor 21 (FGF-21) on the expression of glucose in resistin-overexpressing hepatocytes with insulin resistance HepG2 cells were transfected into HepG2 cells, and HepG2 cells overexpressing resistin were screened by flow cytometry. Cells were stimulated with different concentrations of insulin and FGF-21, respectively, and treated with GOD- POD method was used to detect glucose uptake in cells and the changes of GLUT1 and PPAR-γ mRNA expression after resistin transfection and FGF-21 treatment were detected by real-time fluorescence quantitative PCR.The results of PCR showed that HepG2 overexpressing resistin cells were constructed Successfully.GOD-POD test results showed that model cells decreased insulin sensitivity, but FGF-21 still can effectively regulate the glucose uptake of model cells in a dose-dependent manner.Real-time quantitative real-time PCR assay showed that resistin transfection The expression of GLUT1 mRNA in HepG2 cells was increased. Compared with the control cells, the expression of GLUT1 mRNA was still up-regulated in the model cells stimulated by FGF-21. The changes of PPAR-γ Significant. These results suggest that over-expression of hepatocyte resistin, decreased insulin sensitivity, insulin resistance, FGF-21 but still effectively regulate its glucose metabolism.