高浓度葡萄糖诱导大鼠腹膜间皮细胞上皮-间叶转化

目的探讨高浓度葡萄糖对大鼠腹膜间皮细胞上皮-间叶转化(EMT)的影响,建立大鼠腹膜间皮细胞EMT模型。方法体外培养的大鼠腹膜间皮细胞经含60、120 mmol/L葡萄糖的M199培养基分别培养48、72 h;以正常M199培养基和含120 mmol/L甘露醇的M199培养基为对照。采用相差显微镜观察细胞形态学改变,应用实时定量PCR法检测间皮细胞E-cad-herin、α-SMA、Collagen I mRNA的表达;应用Western印迹法检测E-cadherin、α-SMA蛋白的表达。结果高浓度葡萄糖(60、120 mmol/L)刺激后,大鼠腹膜间皮细胞由典型的上皮细胞形态逐渐变为梭形及不规则形,类似肌成纤维细胞;大鼠腹膜间皮细胞α-SMA、Collagen I mRNA和α-SMA蛋白表达水平显著增加(P0.05);大鼠腹膜间皮细胞E-cadherin mRNA和E-cad-herin蛋白表达水平显著下降(P0.05)。结论高浓度葡萄糖能上调α-SMA、Collagen I表达和下调E-cadherin表达,高浓度葡萄糖诱导大鼠腹膜间皮细胞EMT。

High glucose induce epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells

Objective To investigate the effects of high glucose on epithelial-to-mesenchymal transition(EMT) of rat peritoneal mesothelial cells.Methods The cultured rat peritoneal mesothelial cells were exposed to culture medium containing different concentrations of glucose(60,120 mmol/L),120 mmol/L mannitol and normal M199 culture medium as control for different time(48 hours and 72 hours).Then phenotype changes of rat peritoneal mesothelial cells were observed by light microscopy;The mRNA level and protein expressions of α-smooth muscle actin(α-SMA),E-cadherin,and collagen I were determined with Real-Time PCR and Western blotting,respectively.Results Phenotype changesin rat peritoneal mesothelial cells were observed in High glucose(60,120 mmol/L) group including loss of classic epithelial phenotype and acquirement the appearance similar to myofibroblast;The mRNA levels of α-SMA,collagen I,and protein expression of α-SMA were significantly higher in high glucose(60,120 mmol/L) group than those in mannitol group and normal culture medium group respectively(P<0.05);The mRNA levels and protein expression of E-cadherin was significantly lower in high glucose(60,120 mmol/L) group than that in mannitol group and normal culture medium group respectively(P<0.05).Conclution High glucose can induce epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells,which is proved by loss of E-cadherin and acquirement of α-SMA and Collagen I.