机械牵拉刺激对真皮成纤维细胞胶原合成影响的初步研究  

Preliminary study on the collagenesis in human dermal fibroblasts upon mechanical stretching

作　　者：李晓荟[1] 雷铁池[1] 罗龙飞[1] 史赢[1] 徐世正[1] 

机构地区：[1]武汉大学人民医院皮肤性病科,湖北武汉430060

出　　处：《临床皮肤科杂志》2016年第7期492-495,共4页Journal of Clinical Dermatology

基　　金：中华医学会欧莱雅资助项目(2014HX0023)

摘　　要：目的:探讨机械牵拉刺激对真皮成纤维细胞胶原合成的直接影响。方法 :自人包皮组织建立原代真皮成纤维细胞培养;将培养的成纤维细胞接种至Falcon细胞培养小室(美国BD公司,货号353092),培养小室底部聚乙烯对苯二甲酸酯(polyethylene terephthalate,PET)膜含有直径3μm的小孔,待成纤维细胞生长至融合,通过重力加压致使PET膜变形,借以机械牵拉PET膜表面附着生长的成纤维细胞。用倒置显微镜观察机械牵拉后成纤维细胞形态学改变;用实时荧光定量PCR(qPCR)和蛋白印迹技术检测机械牵拉对成纤维细胞的Ⅰ型胶原蛋白(COL1A),基质金属蛋白酶-1(MMP1),组织基质金属蛋白酶抑制剂-1(TIMP1),α2整合素(ITGA2)和α平滑肌肌动蛋白(ACTA2)5种基因m RNA及其蛋白表达的影响。结果:机械牵拉组细胞被拉长呈细长梭形,细胞长轴沿应力方向排列,而未牵拉对照组细胞呈梭形,排列无序散乱;qPCR结果显示机械牵拉组细胞COL1A,TIMP1,ITGA2和ACTA2基因mRNA表达增加,分别为未牵拉对照组的1.43、1.83、1.62、2.18倍(P<0.05),而MMP1基因表达无明显变化(P>0.05);蛋白印迹结果显示5种蛋白表达改变与其基因m RNA表达一致。结论 :机械牵拉刺激能改变真皮胶原蛋白合成-降解平衡,增加Ⅰ型胶原蛋白的从头合成。Objective： To investigate the effect of mechanical stretching on collagen synthesis in human dermal fibroblasts （HDFBs）. Methods： Primary HDFBs were from human foreskin tissues. Cultured HDFBs were seeded onto the mieroporous membrane inserts （3.0-1μm pore size, BD Falcon, Cat#：353092） and grew to confluence. Mechanical stretching was achieved by pressing the insert polyethylene terephthalate （PET） membrane, which were firmly attached to the cells, to induced distor- tion. The morphological changes of the stretched HDFBs were visualized using an inverted microscopy. The changes in the expression levels of mRNA and protein of collagen type 1 （COL-l）, matrix metalloproteinase-1 （MMP-1）, tissue inhibitor of metalloproteinase-l（TIMP-1）, integrin α2（ITGA2）, and smooth muscle actin α（ACTA2） in the stretched HDFBs were evalu- ated using a quantitative real-time PCR （qPCR） and western blotting, respectively. Results： The stretched cells appeared a spindle elongated shape arranged in the orientation parallel to the mechanical tension direction. In contrast, the control cells were a spindle shape arranged irregularly, qPCR resuhs showed that the mRNA levels of COL1, TIMP1, ITGA2 and ACTA2 increased by 1.43, 1.83, 1.62, and 2.18 folds, respectively, over the controls（P〈0.05）, while MMP1 expression remained un- changed. Meanwhile, the changes in protein expression were consistent with their mRNA levels. Conclusion： Mechanical stretching balances collagen production/degradation and increases de novo synthesis of collagen type I.

关 键 词：成纤维细胞 机械牵拉 胶原合成 老化 

分 类 号：R751[医药卫生—皮肤病学与性病学]