高糖对体外培养人脐静脉内皮细胞通透性及肌球蛋白轻链磷酸化的影响  

Effects of high-glucose on endothelial permeability and MLC phosphorylation of HUVECs in vitro

作　　者：赵晓燕[1] 张金盈[1] 王小芳[1] 李丹花[1] 金强松 

机构地区：[1]郑州大学第一附属医院心血管内科,郑州450052

出　　处：《郑州大学学报（医学版）》2015年第2期198-201,共4页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:探讨高糖对人脐静脉内皮细胞(HUVECs)通透性及肌球蛋白轻链(MLC)磷酸化的影响。方法:分别以不含、含dnRho A或Y27632的常规M199培养基(对照组、dnRho A组、Y27632组),以及不含、含dnRho A或Y27632的高糖M199培养基(高糖组、高糖+dnRho A组、高糖+Y27632组)培养HUVECs,在transwell系统中使其生长为单层内皮结构,观察该结构葡聚糖跨内皮转移率和THP-1跨内皮迁移细胞数,测定细胞MLC磷酸化水平。结果:高糖组葡聚糖跨内皮转移率和THP-1跨内皮迁移细胞数较对照组增加(P<0.05)。dnRho A组、Y27632组和高糖+dnRho A组、高糖+Y27632组葡聚糖跨内皮转移率、THP-1跨内皮迁移细胞数及MLC磷酸化水平均较高糖组降低(P均<0.05)。结论:高糖刺激可能通过激活Rho A-ROCK信号转导通路,上调MLC磷酸化水平,增加内皮通透性。Aim： To observe the effects of high-glucose on endothelial permeability and to explore the mechanism. Methods： HUVECs were cultured with M199 medium（ control group） ,M199 ＋ dnRhoA（ dnRhoA group） , M199 ＋ Y27632 （ Y27632 group） and high-glucose-M199 （ high-glucose group） , high-glucose-M199 ＋ dnRhoA （ high-glucose ＋ dnRhoA group）, high-glucose-M199 ＋ Y27632（ high-glucose ＋ Y27632 group） in transwell system. Dextran transflux and THP-1 transmigration were detected with transwell system, and myosin light chain（MLC） protein phosphorylation was measured by Western blot. Results： Compared with those of the control group, dextran transflux and THP-1 transmigration of high-glucose group increased （P 〈 0.05 ）. Compared with high-glucose group, the dextran transflux, THP-1 transmigration and MLC phosphorylation of dnRhoA group, Y27632 group, high-glucose ＋ dnRhoA group and high-glucose ＋ Y27632 group decreased（ P 〈 0.05 ）. Conclusion： High-glucose might induce endothelial dysfunction through activating RhoA-ROCK pathway which results in upregulating MLC phosphorylation.

关 键 词：人脐静脉内皮细胞 通透性 肌球蛋白轻链 高糖 RHO A-ROCK通路 

分 类 号：R543[医药卫生—心血管疾病]