论文部分内容阅读
目的明确邻苯二甲酸二(2-乙基己)酯(DEHP)染毒对Hep G2细胞胰岛素信号通路活性的影响,预测DEHP诱导人类代谢相关疾病(如2型糖尿病和肥胖)的风险。方法对Hep G2细胞进行持续24 h的DEHP染毒实验,然后回收细胞并分别提取其蛋白和核酸,最后分别通过Western Blot和荧光定量PCR检测胰岛素信号通路关键蛋白和基因的表达情况。结果 DEHP暴露后,Hep G2细胞胰岛素信号通路关键蛋白磷酸化胰岛素受体(p-IR)和磷酸化蛋白激酶B(p-AKT)的表达量在10、50、100、500、1 000μmol/L的DEHP作用下均显著减弱,仅5μmol/L组变化不显著。而IR,AKT和GSK3β的基因和总蛋白表达水平在各个染毒组和对照组并无显著差异。结论 DEHP可诱导Hep G2细胞产生剂量依赖式的胰岛素抵抗,这种效应可能是DEHP增加人类2型糖尿病和肥胖发病风险的潜在分子机制之一。DEHP诱导Hep G2细胞出现胰岛素抵抗的效应可能是发生在蛋白质翻译后修饰水平,而非转录或翻译水平。
Objective To determine the effect of DEHP on the insulin signaling pathway activity of Hep G2 cells and to predict the risk of DEHP-induced human metabolic-related diseases such as type 2 diabetes and obesity. Methods Hep G2 cells were exposed to DEHP for 24 hours. Cells were harvested and their proteins and nucleic acids were extracted. The expression of key proteins and genes in insulin signaling pathway were detected by Western Blot and real-time PCR respectively. Results After DEHP exposure, the expressions of phosphorylated insulin receptor (p-IR) and phosphorylated protein kinase B (p-AKT), key proteins of insulin signaling pathway in Hep G2 cells, were significantly different between 10, 50, 100, 500 and 1000 μmol / L DEHP significantly weakened, only 5μmol / L group change was not significant. However, the expression levels of IR, AKT and GSK3β in all the infected and control groups showed no significant difference. Conclusion DEHP can induce a dose-dependent insulin resistance in Hep G2 cells. This effect may be one of the potential molecular mechanisms by which DEHP can increase the risk of type 2 diabetes and obesity in humans. The effect of DEHP on insulin resistance in Hep G2 cells may be at the level of post-translational modification but not transcription or translation.