7-二氟甲氧基-5,4'-二甲氧基金雀异黄素上调miR-26b对妊娠期糖尿病脂肪细胞葡萄糖摄取的影响

Effects of 7-Difluoromethoxy-5,4'-dimethoxygenistein on Adipocyte Cells of Gestational Diabetes Mellitus In-taking Glucose via Up-regulation miR-26b

关键词:  7-二氟甲氧基-5,4'-二甲氧基金雀异黄素  miR-26b  妊娠期糖尿病

Key words:  7-difluoromethoxy-5,4'-dimethoxygenistein  miR-26b  gestational diabetes mellitus

摘要:

目的 探究7-二氟甲氧基-5，4'-二甲氧基金雀异黄素（7-difluoromethoxy-5，4'-dimethoxygenistein，DFMG）对妊娠期糖尿病（gestational diabetes mellitus，GDM）肠系膜脂肪细胞摄取葡萄糖的影响。方法 提取GDM孕妇肠系膜脂肪细胞，MTT检测不同浓度DFMG对脂肪细胞的毒性；液体闪烁仪检测脂肪细胞对[³H]-2-脱氧-D-葡萄糖的摄取率；RT-PCR检测孕妇肠系膜脂肪细胞miR-26b及其下游靶基因PTEN mRNA表达变化；Western blotting检测IRS-PI3K/AKT-mGLUT4信号通路相关蛋白的表达。结果 DFMG对体外培养GDM孕妇肠系膜脂肪细胞无毒副作用，与GDM组相比，液体闪烁仪提示DFMG可显著促进GDM肠系膜脂肪细胞对[³H]-2-脱氧-D-葡萄糖的摄取率（P<0.05）。RT-PCR提示DFMG可以显著上调GDM脂肪细胞miR-26b表达（P<0.05），下调PTEN mRNA表达（P<0.05）。Western blotting提示DFMG可以下调PTEN蛋白表达（P<0.05），上调p-PI3K、p-AKT和mGLUT4蛋白表达（P<0.05）。结论 DFMG可能通过上调miR-26b表达，调节IRS-PI3K/AKT-GLUT4信号通路，调节GDM肠系膜脂肪细胞对葡萄糖的摄取率。

Abstract:

OBJECTIVE To investigate the effects of 7-difluoromethoxy-5,4'-dimethoxygenistein(DFMG) on adipocyte cells of gestational diabetes mellitus(GDM) in-taking glucose. METHODS Adipocyte cells were extracted from mesenteric adipose tissue of GDM patients. The toxicity of different concentrations of DFMG for adipocyte cells were determined by MTT assay. The rate of adipocyte cells in-taking[³H]-2-decoxy-glucose was evaluated by liquid scintillation detector. The expression of miR-26b and its downstream target gene PTEN mRNA were examined by RT-PCR assay. The proteins expression related to IRS-PI3K/AKT-mGLUT4 signaling pathways were assessed using Western blotting method. RESULTS There was no toxide side effect of DFMG on akipocyte cells of GDM in vitro. Liquid scintillation detector showed DFMG could prompt the rate of adipocyte cells in-taking[³H]-2-decoxy-glucosecompared with GDM group(P<0.05). RT-PCR method indicated DFMG could up-regulate the expression of miR-26b and down-regulate its target gene PTEN mRNA compared with GDM group(P<0.05). Western blotting assay showed DFMG could reduce the expression of PTEN protein compared with GDM group(P<0.05), enhance the expressions of p-PI3K, p-AKT and mGLUT4 proteins(P<0.05). CONCLUSION DFMG can promote the rate of adipocyte cells in-taking[³H]-2-decoxy-glucose via up-regulating miR-26b expression, adjusting the IRS-PI3K/AKT-GLUT4 signaling pathways.