高糖和茶多酚对人脐静脉内皮细胞基质金属蛋白酶2及其诱导因子表达的影响

High glucose and tea polyphenols on expression of matrix metalloproteinase-2 and extracellular matrix metailoproteinase inducer in human umbilical vein endothelial cells

目的探讨高糖对内皮细胞基质金属蛋白酶2（MMP-2）及其诱导因子（EMMPRIN）表达的影响，以及茶多酚对高糖作用的干预情况。方法培养人脐静脉内皮细胞，分为正常对照组、高糖组、茶多酚对照组和茶多酚干预组。培养24h后，用免疫细胞化学、RT-PCR和Western blot法测定细胞内MMP-2及EMMPRIN的变化。结果高糖下调内皮细胞MMP-2及EMMPRIN表达，其变化随时问延长更为明显；而茶多酚干预组MMP-2活性明显上调（P〈0．05）；茶多酚干预组EMMPRIN mRNA及蛋白表达均较高糖组上调（P〈0．05），茶多酚干预可拮抗高糖对内皮细胞的影响。结论高糖影响内皮细胞的细胞外基质降解，茶多酚可以抑制这种作用。

Objective To investigate the effects of high glucose on expression of matrix metalloproteinase-2 （MMP-2） and extracellular matrix metalloproteinase inducer （EMMPRIN） in human umbilical vein endothelial cells （HUVECs）. Methods HUVECs were cultured in DMEM media containing high glucose with or without polyphenols for 24 hours respectively. The activity of MMP-2 in the supernatant was detected by zymography. The expression of MMP-2 mRNA and protein in HUVECs were detected by RTPCR and Western blot respectively. The expression of EMMPRIN mRNA and protein in the cells were determined by RT-PCR as well as immunocytochemistry and Western blot respectively. Results The expression of MMP-2 and EMMPRIN mRNA were suppressed by a high concentration of glucose. Both the MMP-2 activity and protein level of EMMPRIN expression were also significantly decreased. Polyphenols abolished all the above changes of HUVECs induced by a high glucose level （P 〈 0. 05 ）. Conclusions An acute high glucose stimulation down-regulates the activity of MMP-2, the expressions of MMP-2 and EMMPRIN at RNA and protein levels in the endothelial cells, which may play an important roles in diabetic vascular complications in the early phase. Polyphenols treatment can diminish the detrimental effects of high glucose on HUVECs.