高糖高脂促进人脐静脉内皮细胞间充质转化

High Glucose and Fat Promoted Mesenchymal Transformationof Human Umbilical Vein Endothelial Cells

目的探究高糖高脂刺激对人脐静脉内皮细胞(HUVEC)间充质转化的影响。方法将培养至第6代的HUVEC设为对照组(CON组),以常用的间充质转化诱导剂TGF-β1为阳性对照(TGF-β1组),给予33mmol/L高糖加0.1mmol/L棕榈酸模拟高糖高脂环境对HUVEC进行刺激为HG+HP组,显微镜下观察细胞形态变化。并通过免疫荧光、Western-blot法检测内皮细胞特异性的标记因子VE-Cadherin和CD31,以及平滑肌细胞特异性标记因子α-SMA和N-Cadherin等蛋白的表达情况。结果显微镜下观察发现,HUVEC经过TGFβ1培养处理72h后,细胞形态由饱满圆润的短梭形逐渐变成了长梭形,细胞间隙变大,与CON组相比有明显改变,而HG+HP组细胞形态与TGF-β1组相似。免疫荧光和Western-blot结果显示,与CON组相比,HG+HP组内皮细胞标记物VE-Cadherin和CD31表达量显著下降(P<0.05),而平滑肌细胞标记物α-SMA和N-Cadherin表达量显著上升(P<0.05)。结论高糖高脂刺激可以促进人脐静脉内皮细胞的间充质转化。

Objective To investigate the effect of high sugar and high fat stimulation on mesenchymal transition of human umbilical vein endothelial cells(HUVEC).Methods HUVEC cultured to the 6th generation were set as the control group.TGF-β1,a commonly used mesenchymal transition inducer,was usingas a positive control.HUVEC were stimulated with 33mmol/L high glucose plus 0.1mmol/L palmitic acid to simulate a high glucose and high lipid environment and set as the HG+HP group.The morphological changes of the cells were observed bythe microscope.The expression of endothelial cell-specific markers VE-Cadherin and CD31,and smooth muscle cell-specific markersα-SMA and N-Cadherin were detected by immunofluorescence and Western-blot.Results Microscopic observation revealed that the cell morphology of HUVEC changed from cobblestone-like to elongatedand intercellular space increased after the 72 h of TGFβ1 culture treatment,and the cell morphology was significantly altered compared to the control group,while the cell morphology of the high glucose and high lipid group was similar to that of the TGFβ1 group.Immunofluorescence and Western-blot results showed a significant decrease of endothelial cell-specific markers VE-Cadherin and CD31 expression(P<0.05)and a significant increase of the smooth muscle cell-specific markersα-SMA and N-Cadherin expression(P<0.05)in the high glucose and high fat group compared to the control group.Conclusion High glucose and high fat stimulation plays an important role in promoting mesenchymal transformation of human umbilical vein endothelial cells.