摘要
目的 探讨不同糖浓度状态下人脐静脉内皮细胞(HUVECs)形态、增殖活性及VE-钙黏蛋白(VE-cadherin)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)表达变化.方法 将HUVECs分为正常对照组(葡萄糖浓度5.5 mmol/L)、甘露醇组(葡萄糖浓度5.5 mmol/L+甘露醇浓度24.5 mmol/L)、高糖组(葡萄糖浓度分别为11.1、22.2、33.3 mmol/L),分别培养24、48、72 h,以显微镜观察细胞形态,用CCK-8法检测各组细胞增殖情况,Western blotting法检测各组HUVECs VE-cadherin、E-cadherin、N-cadherin表达.结果 培养72 h,正常对照组细胞形态呈类圆形,高糖组、甘露醇组细胞形态由圆形、椭圆形逐渐到条索状,细胞数量减少、细胞间隙较前增加.培养24 h,高糖组细胞增殖率高于正常对照组;随糖浓度升高,高糖组细胞增殖率呈现先升高后降低趋势.培养48、72 h,高糖组细胞增殖率低于正常对照组;随着糖浓度升高,高糖组细胞增殖率逐渐降低.以上各组细胞增殖率比较均有统计学差异(P均<0.05).培养24、72 h,与正常对照组比较,高糖组HUVECsN-cadherin、VE-cadherin、E-cadherin蛋白表达明显升高(P均<0.05);且随着葡萄糖浓度升高,高糖组细胞N-cadherin、VE-cadherin、E-caderin蛋白表达呈现先升高后降低的趋势.结论 在高糖条件下,HUVECs发生形态转变,增殖率下降,且伴有细胞钙黏蛋白N-cadherin、VE-cadherin、E-cadherin表达变化.
Abstract
Objective To explore the changes in morphology,proliferation activity and VE-cadherin,E-cadherin,and N-cadherin expression of human umbilical vein endothelial cells(HUVECs)under different concentrations of glucose.Methods HUVECs were divided into the normal control group(glucose concentration of 5.5 mmol/L),mannitol group(glucose concentration of 5.5 mmol/L+mannitol concentration of 24.5 mmol/L),and high glucose group(glucose con-centration of 11.1,22.2,and 33.3 mmol/L,respectively),and were cultured for 24,48,and 72 h.Cell morphology was observed under the microscope,and cell proliferation in each group was detected by CCK-8 reagent.Western blotting was used to detect the expression levels of VE-cadherin,E-cadherin,and N-cadherin in HUVECs in each group.Results After 72 h of cultivation,the cells in the normal control group showed a quasi-circular shape,while cells in the high glucose group and mannitol group gradually changed from circular and elliptical to cord shape,with a decrease in cell number and an increase in cell gaps compared with those before.After 24 h of cultivation,the cell proliferation rate of the high glucose group was higher than that of the normal control group.With the glucose concentration increased,the cell pro-liferation rate of the high glucose group showed a trend of first increasing and then decreasing.After 48 and 72 h of cultiva-tion,the cell proliferation rate of the high glucose group was lower than that of the normal control group.With the glucose concentration increased,the cell proliferation rate of the high glucose group gradually decreased.Statistically significant differences were found in the cell proliferation rates among the above groups(all P<0.05).After 24 and 72 h of cultiva-tion,compared with the normal control group,the expression levels of N-cadherin,VE-cadherin,and E-cadherin in HUVECs increased in the high glucose group(all P<0.05);and with the increase of glucose concentrations,the N-cad-herin,VE-cadherin,and E-cadherin of cells in the high glucose group showed a trend of first increasing and then decreas-ing.Conclusion Under high glucose conditions,HUVECs undergo morphological transformation,with a decrease in proliferation rate,accompanied by changes in the expression levels of N-cadherin,VE-cadherin,and E-cadherin.
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