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目的旨在探讨体外培养条件下高糖状态对血管内皮细胞的影响。方法以原代培养人脐带静脉内皮细胞为材料 ,用Fura 2荧光检测法观察高糖对原代培养血管内皮细胞内钙离子浓度([Ca2 + ]i)的影响 ,用放射免疫分析法观察高糖作用下血管内皮细胞分泌基底膜蛋白成分IV型胶原蛋白 (IV C)和层粘连蛋白 (LN)的情况。结果在 1 1 1 1mmol L、1 6 6 7mmol L和 33 33mmol L糖浓度下培养 2 4小时 ,血管内皮细胞 [Ca2 + ]i分别升高了 1 74、2 84和 6 74倍。发现不同糖浓度作用下血管内皮细胞分泌IV C的量均无变化 ;在 1 6 6 7mmol L糖浓度下培养 2 4小时血管内皮细胞分泌LN的量下降了 2 6 6 9%。结论在较短时间内糖浓度升高可引起血管内皮细胞 [Ca2 + ]i升高 ,而对血管内皮细胞分泌LN起抑制作用 ,对IV C的分泌无明显影响。在糖尿病微血管病变早期 ,高糖可能通过血管内皮细胞 [Ca2 + ]i升高而起主要作用
The aim of this study was to investigate the effect of high glucose on vascular endothelial cells under in vitro culture conditions. Methods Primary cultured human umbilical vein endothelial cells were used as materials. The effect of high glucose on the intracellular Ca2 + concentration ([Ca2 +] i) in primary cultured human umbilical vein endothelial cells was observed by Fura2 fluorescence assay. The radioimmunoassay Vascular endothelial cells secrete basement membrane protein components type IV collagen (IV C) and laminin (LN) under the action of glucose. The results showed that [Ca2 +] i increased by 1 74, 2 84 and 6 74 times, respectively, after cultured for 24 hours with 1 1 1 1 mmol L, 1 6 6 7 mmol L and 33 33 mmol L glucose. It was found that there was no change in the amount of IVC secreted by vascular endothelial cells under different concentrations of glucose. The amount of LN secreted by vascular endothelial cells decreased by 269% after culturing for 24 hours at a concentration of 1 667 mmol L glucose. Conclusion The increase of glucose concentration in a short period of time can cause the increase of [Ca2 +] i in vascular endothelial cells and inhibit the secretion of LN in vascular endothelial cells, but has no obvious effect on IVC secretion. In the early stage of diabetic microangiopathy, high glucose may play a major role through the increase of [Ca2 +] i in vascular endothelial cells