人体不同部位正常皮肤成纤维细胞对机械张力反应的研究  

REACTION OF HUMAN FIBROBL ASTS FROM DIFFERENT SITES TO THE MECHANICAL STRESS

作　　者：王志国[1] 匡瑞霞[1] 徐全臣[2] 陈振雨[1] 李鹏[3] 

机构地区：[1]青岛大学附属医院烧伤整形外科,青岛266003 [2]青岛大学附属医院口腔科,青岛266003 [3]潍坊市人民医院整形外科

出　　处：《中国修复重建外科杂志》2015年第4期467-471,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：山东省自然科学基金资助项目(ZR2013HQ006);青岛市应用基础研究项目(13-1-4-161-jch)~~

摘　　要：目的探讨人体不同部位正常皮肤成纤维细胞对机械张力反应的差异。方法取色素痣切除患者自愿捐赠的背部和上臂内侧正常皮肤,应用组织块法体外培养成纤维细胞,取第5～8代细胞进行实验。将背部和上臂内侧正常成纤维细胞分别设实验组和对照组,实验组细胞采用多通道细胞应力加载仪加载周期性机械张力24、36、48 h;对照组细胞正常培养。各时间点加载结束后,倒置显微镜下观察细胞形态变化;CCK-8法检测细胞增殖活性;RT-PCR法检测细胞内整合素β1、P130Ca s（p130Crk-associated substance）、TGF-β1、Ⅰ型胶原a1链（collagen type Iα1 chain,COL1A1）m RNA水平;ELISA法检测Ⅰ型胶原和TGF-β1含量。对照组于培养对应时间取细胞进行以上观察。结果实验组加载后细胞均增殖旺盛,分布密集,排列呈一定方向性。加载24 h后,实验组背部及上臂内侧细胞增殖,整合素β1、P130Cas、TGF-β1 m RNA表达水平和TGF-β1含量比较,差异均无统计学意义（P〉0.05）;加载36、48 h时,背部细胞以上检测指标均显著高于上臂内侧细胞,差异有统计学意义（P〈0.05）。实验组各时间点背部及上臂内侧细胞的COL1A1 m RNA表达水平和Ⅰ型胶原含量比较,差异均无统计学意义（P〉0.05）。对照组各时间点背部及上臂内侧细胞以上指标比较,差异均无统计学意义（P〉0.05）。结论背部和上臂内侧皮肤成纤维细胞对机械张力的反应不同,提示人体皮肤成纤维细胞的力学特征存在部位差异,可能导致不同部位增生性瘢痕的发生率不同。Objective To explore the reaction of normal skin fibroblasts from different sites of human body to cyclic stretch. Methods The normal skin tissues from scapular upper back and medial side of upper arm of 3 patients were cultured in vitro. Fibroblasts of experimental group were loaded by cyclic stretch with 10% amplitude for 24, 36, and 48 hours respectively. Fibroblasts of control group were cultured without cyclic stretch. The morphologic changes were observed using inverted microscope. CCK-8 method was used to detect the proliferation of the fibroblasts. The expressions of integrin β1 m RNA, p130Crk-associated substance（P130Cas） m RNA, transform growth factor β1（TGF-β1） m RNA, and collagen type I α1 chain（COL1A1） m RNA were detected by real-time quantitative PCR. The protein levels of collagen type I and TGF-β1 were detected by ELISA. Results The cultured cells showed a significantly increased cell proliferation ability, and apparent orientation after the applied strain. The proliferation activity, mRNA expression levels of integrin β1, P130 Cas, and TGF-β1, protein levels of TGF-β1 in back skin were significantly higher than those in arm skin（P〈0.05） when the fibroblasts were loaded for 36 and 48 hours, but no significant difference between back skin and arm skin at 24 hours（P〉0.05）. There was no significant difference in m RNA expression level of COL1A1 and protein level of collagen type I between back skin and arm skin at 24, 36, and 48 hours（P〉0.05）. There was no significant difference in all above indexes between back skin and arm skin in control group（P〉0.05）. Conclusion Fibroblasts from scapular upper back and medial side of upper arm display different reactions to cyclic stretch, which indicates that there exists site difference in the reactions of fibroblasts to cyclic stretch. It might be related with the incidence of hypertrophic scar in different sites of the body.

关 键 词：成纤维细胞 机械张力 正常皮肤 增生性瘢痕 

分 类 号：R622[医药卫生—整形外科]