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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 m RNA表达变化;Western blotting检测IRS-PI3K/AKT-m GLUT4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mi R-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。
Objective To investigate the effect of 7-difluoromethoxy-5,4’-dimethoxygenistein (DFMG) on mesenteric fat in gestational diabetes mellitus (GDM) The effect of glucose uptake by cells. Methods Mesenteric adipocytes of GDM pregnant women were extracted and MTT was used to detect the cytotoxicity of DFMG on adipocytes. The uptake of [3H] -2-deoxy-D-glucose by adipocytes was detected by liquid scintillant. -26b and its downstream target gene PTEN m RNA expression; Western blotting detected IRS-PI3K / AKT-m GLUT4 signal pathway related protein expression. Results Compared with GDM group, DFMG had no toxic side effects on mesentery adipose cells cultured with GDM in vitro, and liquid scintillator suggested that DFMG could significantly promote the uptake of [~ 3H] -2-deoxy-D-glucose by GDM mesentery adipocytes <0.05). RT-PCR indicated that DFMG could upregulate the expression of mi R-26b (P <0.05) and down-regulate the expression of PTEN mRNA in GDM adipocytes (P <0.05). Western blotting suggested that DFMG could down-regulate the expression of PTEN protein (P <0.05) and up-regulate the expression of p-PI3K, p-AKT and mGLUT4 protein (P <0.05). Conclusion DFMG may up regulate the expression of miR-26b and regulate IRS-PI3K / AKT-GLUT4 signaling pathway to regulate glucose uptake in GDM mesenteric adipocytes.