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目的:观察高浓度葡萄糖诱导人晶状体上皮细胞发生上皮-间质转分化(epithelial-to-mesenchymal transition,EMT)。方法:将人晶状体上皮细胞HLE-B3系分别培养在正常葡萄糖浓度(5.5 mmol/L)DMEM培养基和高浓度葡萄糖(35.5 mmol/L)的DMEM培养基中24小时,于培养的0 h、3 h、6 h、12 h、24 h在倒置显微镜下观察细胞形态学变化,采用免疫荧光染色检测晶状体上皮细胞中EMT相关蛋白E-cadherin及α-SMA的表达变化。结果:与正常糖浓度组相比,随着时间的延长高糖组细胞逐渐丢失上皮细胞形态,细胞变细、变长,向纤维细胞的形态转变;同时随着时间的延长,高糖组晶状体上皮细胞中E-cadherin染色的荧光强度在各时间点均低于正常糖浓度组,而α-SMA的荧光强度却明显高于正常糖浓度组,在6 h和12 h时差异显著,有统计学意义(P<0.01)。结论:高浓度葡萄糖诱导人晶状体上皮细胞发生上皮-间质转分化。
Objective: To observe the epithelial-to-mesenchymal transition (EMT) induced by high glucose in human lens epithelial cells. Methods: Human lens epithelial cells (HLE-B3) were cultured in DMEM medium with normal glucose concentration (5.5 mmol / L) and high concentration glucose (35.5 mmol / L) for 24 hours respectively. 3 h, 6 h, 12 h, 24 h under inverted microscope. The expression of EMT-related protein E-cadherin and α-SMA in lens epithelial cells were detected by immunofluorescence staining. RESULTS: Compared with normal glucose concentration group, the morphology of epithelial cells was gradually lost in high glucose group as time went by, the cells became thinner and longer, and the morphology of fibroblasts changed. At the same time, with the prolongation of time, The fluorescence intensity of E-cadherin staining in epithelial cells was lower than normal glucose concentration group at each time point, while the fluorescence intensity of α-SMA was significantly higher than that of normal glucose concentration group, and the difference was significant at 6 h and 12 h Significance (P <0.01). Conclusion: High concentration glucose induces epithelial-mesenchymal transition in human lens epithelial cells.