缺氧对创面修复主要细胞MMP.doc

低氧环境对创面修复主要效应细胞表达MMP-9的作用研究 黄翀，黄跃生，张家平，江旭品，张琼，贾丽杰 （第三军医大学西南医院全军烧伤研究所，创伤、烧伤、复合伤国家重点实验室，重庆 400038） [摘要] 目的 探讨低氧（2%氧气）对创面修复主要效应细胞（表皮细胞及真皮成纤维细胞）表达MMP-9的影响。方法 常规培养人表皮细胞及人真皮成纤维细胞，分为常氧组和低氧组（设低氧培养3h和6h两个组）。分别采用Western blot、明胶酶谱法和荧光定量PCR法检测人表皮细胞和真皮成纤维细胞MMP-9蛋白表达、 MMP-9胞外分泌和MMP-9转录水平变化。结果 在常氧培养的表皮细胞中，MMP-9呈一定程度的基础表达和胞外分泌；低氧能明显促进表皮细胞MMP-9的蛋白表达（6h时增强约20%，与常氧组比较，P＜0.01）和MMP-9的胞外分泌（常氧组：44.03±3.00；低氧6h组： 60.64±3.77。二者比较，P＜0.01）。相反，无论是在常氧或低氧培养条件下，真皮成纤维细胞几乎都不表达和分泌MMP-9。荧光实时定量 PCR结果显示，低氧培养条件下表皮细胞MMP-9 的mRNA水平显著增高：常氧对照组为1.67±0.692，低氧6h组为5.47±0.507，P=0.002。结论 低氧刺激能显著诱导表皮细胞表达和分泌MMP-9，这一作用可能与其促进MMP-9的转录有关。研究提示表皮细胞，而非真皮成纤维细胞，是低氧环境诱导创面局部组织表达MMP-9的重要效应细胞。 [关键词] 创面愈合；低氧；HaCaT细胞；人成纤维细胞；MMP-9。 [中图法分类号] R322 991；R345；R329 28 [文献标识码] A 基金项目：1) 国家自然科学基金面上项目； 2）国家“973”01课题分题（2012CB518101） 通讯作者：黄跃生，E-mail：yshuang.tmmu@ Expression of MMP-9 in the main effector cells in wound repair infuenced by hypoxia. Huang chong;Huang yuesheng;Zhang jiaping;Jiang xupin;Zhang qiong;Jia lijie. (Institute of Burn Research Southwest Hospital Third Military Medical University Chongqing, Abstract Objective To discuss the effect of MMP-9 expression of main effector cells(epidermal cells and fibroblasts) in wound repair influenced by hypoxia(2% oxygen).Methods Routinely culture the human epidermal cells and fibroblasts and divide them into two groups: normoxic group (synchronous culture, no treatment), hypoxia group (two subgroups of 3hrs and 6hrs under hypoxia).Detect the MMP-9 expressions of epidermal cells and fibroblasts、extracellular secretion of MMP-9 precursor expressions and changes of MMP-9 under the level of transcription with the methods respectively of Western blot, gelatin zymography and fluorescence quantitative PCR. Results MMP-9 has a certain degree of basal expression and extracellular secretion of epidermal cells in normoxic group. Hypoxia can significantly promote the expression levels of MMP-9 protein in the epidermal cells ( 20% increased at 6h compared with the normoxic group,P