机械牵张对兔角膜成纤维细胞细胞外基质基因表达的影响  

Effects of mechanical stretch on gene expression of extracellular matrix in rabbit corneal fibroblasts

作　　者：刘成星[1,2] 冯鹏飞[1] 李晓娜[1] 宋婕[1] 安美文[1] 陈维毅[1] 

机构地区：[1]太原理工大学应用力学与生物医学工程研究所,太原030024 [2]山西医科大学基础医学院,太原030001

出　　处：《医用生物力学》2014年第5期447-453,458,共8页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(11032008;31271005);山西省自然科学基金资助项目(2012011046-2)

摘　　要：目的研究机械牵张与白介素-1β(IL-1β)联合作用对兔角膜成纤维细胞细胞外基质相关基因表达的影响。方法对原代提取的兔角膜成纤维细胞进行牵张幅度15%、频率0.1 Hz的周期性牵张12、24、36 h,同时给予IL-1β处理,采用实时荧光定量PCR检测细胞基质金属蛋白酶(MMPs)、基质金属蛋白酶的组织抑制剂(TIMP-1)和I型胶原α1(Collagen Iα1)mRNA表达变化水平。结果 IL-1β单独作用可以诱导角膜成纤维细胞MMP-1、MMP-3和MMP-9 mRNA表达;MMP-1和MMP-3 mRNA表达随时间而降低,MMP-9、TIMP-1、Collagen Iα1 mRNA则随时间而增加;IL-1β与机械牵张联合作用使MMP-1、MMP-3、MMP-9 mRNA表达水平上调,TIMP-1和Collagen Iα1 mRNA表达下调,且具有时间依赖。单独机械牵张使Collagen Iα1 mRNA表达下降,IL-1β与机械牵张联合作用使其表达进一步下调,且具有时间依赖性。结论机械牵张与炎性因子联合作用可加剧角膜组织破坏,促进角膜膨隆的发生发展。Objective To explore the combined effects of mechanical stretch and interleukin-1β （IL-1β） on gene expression of extracellular matrix in rabbit corneal fibroblasts.Methods Isolated rabbit corneal fibroblasts were subjected to 15％ equibiaxial stretch at frequency of 0.1 Hz for 12 h,24 h and 36 h,respectively,in presence of IL-1β.The gene expressions of matrix metalloproteinases （MMPs）,tissue inhibitor of metalloproteinases 1 （TIMP-1） and collagen type Ⅰ alpha 1 （Collagen Ⅰα1） were detected by real-time quantitative PCR.Results The mRNA levels of MMP-1,MMP-3 and MMP-9 could be up-regulated by IL-1β alone.However,MMP-1 and MMP-3 mRNA levels decreased with time,while MMP-9,TIMP-1 and collagen Ⅰα1 increased with time.Compared with corresponding IL-1β treatment with mechanical stretch groups,the mRNA levels of MMP-1,MMP-3 and MMP-9 were increased and the mRNA levels of TIMP-1 and collagen Ⅰα1 were decreased in a time-dependent manner.The mRNA level of Collagen Ⅰα1 was decreased by loading mechanical stretch alone,and would further decrease time-dependently in combination with IL-1β treatment.Conclusions Mechanical stretch combined with IL-1β may facilitate the corneal tissue damage,thereby contribute to the development of keratectasia.

关 键 词：周期性牵张 基质金属蛋白酶 胶原 角膜成纤维细胞 

分 类 号：R318.01[医药卫生—生物医学工程]