机械牵张力加载时间对诱导正常皮肤成纤维细胞向增生性瘢痕成纤维细胞转化过程的影响研究

The Study of the Effect of Mechanical Stretch Loading Time on the Transformation of Normal Skin Fibroblasts to Hypertrophic Scar Fibroblasts

目的:探究机械牵张力不同加载时间对诱导正常皮肤成纤维细胞(Normal skin fibroblasts,NSFB)向增生性瘢痕成纤维细胞(Hypertrophic scar fibroblasts,HSFB)转化的影响。方法:同一人体NSFB和HSFB分别设实验组和对照组,实验组加载0.1Hz、10%拉伸幅度的机械牵张力3d、5d、7d,对照组细胞均不加力。CCK-8法检测细胞增殖能力;SYBR Green q PCR法检测细胞p130Cas、Integrin-β1、转化生长因子-β_1(TGF-β_1)、Ⅰ型胶原、α-平滑肌肌动蛋白(α-SMA)m RNA表达水平;ELISA法检测细胞上清液TGF-β_1和Ⅰ型胶原含量;Western-blot法检测细胞Integrin-β_1、p130Cas、α-SMA蛋白表达水平。结果:机械牵张力加载5d时,NSFB实验组与HSFB对照组比较,细胞增殖水平及p130Cas、Integrin-β_1、TGF-β_1、Ⅰ型胶原、α-SMA表达水平相近,差异均无统计学意义(P>0.05);加载3d、7d时,以上指标检测结果差异显著,差异均有统计学意义(P<0.05)。结论:0.1Hz、10%拉伸幅度的机械牵张力加载5d可使NSFB表现出HSFB的部分生化特征,机械牵张力对NSFB向HSFB转化过程有明显的诱导作用。

Objective To investigate the effect of mechanical stretch at different loading times on the transformation of normal skin fibroblasts（NSFB） into hypertrophic scar fibroblasts（HSFB）. Methods The same demic NSFB and HSFB were respectively divided into the experimental groups and the control groups. The experimental groups were loaded with 0.1 Hz, 10% amplitude cyclic stretch 3, 5 and 7 days. The control groups were cultured without cyclic stretch. CCK-8 method was used to measure the cell proliferation. SYBR Green q PCR method was used to measure the expression of p130 Crk-associated substance（p130 Cas）, Integrin-β1 transforming growth factor β1TGF-β1, type I collagen and Alpha-smooth muscle actin（α-SMA）. ELISA method was used to measure the content of type I collagen and TGF-β1in culture supernatant. Western-Blot method was used to detect the content of protein Integrin-β1 p130 Cas and α-SMA. Results With 5-days-long amplitude cyclic stretch, cell proliferation and expression of p130 Cas, Integrin-β1 TGF-β1 α-SMA and type I collagen of the NSFB experiment group showed no significant difference with that of the HSFB control group（P〉0.05）. With 3-days-long and 7-days-long amplitude cyclic stretch, the above indexes showed significant differences（P〈0.05）. Conclusion The mechanical tension loading of 0.1 Hz and 10% tensile amplitude 5 d can make NSFB exhibit some biochemical characteristics of HSFB, and the mechanical tension can obviously induce the transformation of NSFB to HSFB.