高糖对大鼠血管平滑肌细胞迁移的影响及其机制  

Effect of High Glucose on the Migration of Vascular Smooth Muscle Cells and Its Mechanisms

作　　者：楚海荣[1] 李宏[1] 吴海燕[1] 苏绍娟[1] 张静[1] 刘建华[1] 成敏[1] 

机构地区：[1]潍坊医学院医学研究中心,山东省潍坊市261053

出　　处：《中国动脉硬化杂志》2014年第11期1097-1100,共4页Chinese Journal of Arteriosclerosis

基　　金：山东省自然科学基金项目(ZR2013CQ032);山东省高等学校科技计划项目(J11LF17);山东省中医药科技发展计划项目(2013-239);潍坊市科技发展计划项目(20121228)

摘　　要：目的探讨高糖对大鼠血管平滑肌细胞（VSMC）迁移的影响及机制。方法利用组织块贴壁法培养大鼠主动脉VSMC,以3~5代细胞为靶细胞,待细胞融合后用2%胎牛血清的DMEM同步化12 h,再经含低糖（5.5mmol/L葡萄糖）、高糖（25 mmol/L葡萄糖）及甘露醇（5.5 mmol/L葡萄糖＋19.5 mmol/L甘露醇）的DMEM处理24h。以改良Boyden小室观察VSMC迁移能力的变化,细胞免疫荧光分析骨架蛋白F-actin的改变,荧光定量RT-PCR分析收缩型平滑肌标志基因α-SMA和合成型平滑肌标志基因骨桥蛋白（OPN）以及基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）基因的表达情况。结果高糖促进VSMC迁移。在高糖环境下,VSMC由收缩型向合成型转变,即α-SMA的表达量明显下降,而OPN的表达量明显升高;高糖促进MMP-2、MMP-9的基因表达（分别为低糖组的3.12倍和2.22倍）,使F-actin的排列发生明显改变。结论高糖促进VSMC迁移,其可能的机制涉及VSMC的表型转变、基质金属蛋白酶表达及F-actin排列的改变。Aim To investigate the effects and mechanisms of high glucose on the migration of vascular smooth muscle cells（ VSMC）. Methods VSMC were isolated from rat arteriae aorta. The 3rd ~ 5th VSMC were incubated with low glucose（ 5. 5 mmol / L）,high glucose（ 25 mmol / L） or mannitol（ 5. 5 mmol / L glucose ＋ 19. 5 mmol / L mannitol）. After 24 h,VSMC migration was assayed with modified Boyden chamber. The effects of high glucose on F-actin cytoskeleton were analysed by immunofluorescence technique. The gene expression of α-SMA,osteopontin（ OPN）,MMP-2 and MMP-9 was determined by real time RT-PCR. Results The migration of VSMC was obviously increased when the cells were cultured with high glucose. Compared with the low glucose group,mRNA levels of MMP-2 and MMP-9 increased obviously in the high glucose group. Moreover,high glucose induced VSMC conversion from contractile phenotype to synthetic phenotype and led to the reorganization of cytoskeleton. Conclusion High glucose promotes the migration of VSMC via complex mechanisms,involved in regulating VSMC phenotype transformation,increasing MMP expression and reorganizing cytoskeleton.

关 键 词：高糖 血管平滑肌细胞 表型转化 细胞迁移 

分 类 号：R363[医药卫生—病理学]