高糖对原代培养人血管内皮细胞TGF-β基因表达和分泌LN及C-IV的影响

Influence of high glucose concentration on the expression of TGF-β gene,excretion of LN and colage-IV in HUVEC

本研究旨在探讨体外培养条件下高糖状态对人血管内皮细胞的影响。以原代培养人脐带静脉内皮细胞为材料 ,采用RT PCR技术观察高糖状态下原代培养血管内皮细胞转化生长因子 β(TGF β)基因表达水平 ,用放射免疫分析法观察高糖作用下的血管内皮细胞分泌基底膜蛋白成分IV型胶原蛋白 (IV C)及层粘连蛋白 (LN)的情况。在16 6 7mmol LD 葡萄糖浓度作用 6、12和 2 4h ,血管内皮细胞TGF β基因表达分别增高了 0 38、1 2 2和 1 4 3倍 ,而血管内皮细胞分泌IV C的量无变化 ;在 16 6 7mmol L糖浓度下培养 2 4h血管内皮细胞分泌LN的量下降了 5 5 6 0 %。结果显示在较短时间内糖浓度升高可引起血管内皮细胞TGF β基因表达增强 ,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。在微血管病变早期 ,高糖可能通过血管内皮细胞TGF β基因高表达而起主要作用。

We investigated the influence of high glucose concentration on human umbilical vein endothelial cell(HUVEC).RT-PCR was used to analyzed the expression of transforming growth factor beta(TGF-β) gene in HUVEC. Type IV collegan(IV-C) and laminin(LN) which were secreted into media by HUVEC were determined by radioimmunoanalysis(RIA).The expression of TGF-β gene in HUVEC treated by high glucose concentration(16.67mmol/L D-glucose) for 6, 12 and 24 hours showed a 0.38, 1.22 and 1.43-fold increase as compared with normal control(5.56mmol/L of D-glucose ). However, the concentration of IV-C in media, which was excreted by HUVEC treated by high glucose concentration did not change. The level of LN in media decreased by 55.60% compared as with control.The results suggest that TGF-β may play an important role in the early phase of microvascular complications.