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(1)
[摘要]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 qPCR法检测细胞p130Cas、Integrin-β1、转化生长因子-β1(TGF-β1)、Ⅰ型胶原、α-平滑肌肌动蛋白(α-SMA)mRNA表达水平；ELISA法检测细胞上清液TGF-β1和Ⅰ型胶原含量；Western-blot法检测细胞Integrin-β1、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-β1、TGF-β1、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。
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[关键词]增生性瘢痕；机械张力；成纤维细胞；细胞转化
[中图分类号]R619+.6 [文献标志码]A [文章编号]1008-6455(2018)06-0055-05
Abstract： 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 0.1Hz， 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 qPCR method was used to measure the expression of p130Crk-associated substance (p130Cas) ， Integrin-β1， transforming growth factor β1 (TGF-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-β1 in culture supernatant. Western-Blot method was used to detect the content of protein Integrin-β1， p130Cas and α-SMA. Results With 5-days-long amplitude cyclic stretch， cell proliferation and expression of p130Cas， 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 0.1Hz and 10% tensile amplitude 5d can make NSFB exhibit some biochemical characteristics of HSFB， and the mechanical tension can obviously induce the transformation of NSFB to HSFB.
, 百拇医药
Key words： hypertrophic scar； mechanical stress； fibroblasts； cell transformation
皮膚损伤后，创面周围的正常皮肤成纤维细胞(Normal skin fibroblasts， NSFB)增殖迁移，分泌转化生长因子-β(TGF-β)使NSFB向增生性瘢痕成纤维细胞(Hypertrophic scar fibroblasts， HSFB)转化，合成以Ⅰ/Ⅲ型胶原蛋白为主的细胞外基质修复组织，并分泌α-平滑肌肌动蛋白(α-SMA)使组织内纤维收缩，导致增生性瘢痕形成。临床研究发现，切口张力与增生性瘢痕形成呈正相关，表明机械张力是导致增生性瘢痕形成的重要因素[1-2]。实验研究发现，加载机械牵张力后的成纤维细胞有表现出HSFB某些表型的趋势。但是，由于缺乏相应增生性瘢痕成纤维细胞模型，机械张力诱导增生性瘢痕形成的具体机制尚未明确。前期研究表明，加载10%拉伸幅度的周期性机械张力时，NSFB会产生明显的机械传导效应和生化反应[3]，但加载时间对诱导NSFB向HSFB转化过程的影响尚未明确。本实验拟采用机械牵张力诱导NSFB向HSFB转化，观察不同机械牵张力加载时间对NSFB中各生化因子表达水平的影响，探究机械牵张力诱导NSFB向HSFB转化的最佳加载时间，为增生性瘢痕形成机制研究进程奠定前期必要基础，并提供重要实验支撑。, 百拇医药(朱明宇 李雨珊 王志国)
, http://www.100md.com
[关键词]增生性瘢痕；机械张力；成纤维细胞；细胞转化
[中图分类号]R619+.6 [文献标志码]A [文章编号]1008-6455(2018)06-0055-05
Abstract： 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 0.1Hz， 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 qPCR method was used to measure the expression of p130Crk-associated substance (p130Cas) ， Integrin-β1， transforming growth factor β1 (TGF-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-β1 in culture supernatant. Western-Blot method was used to detect the content of protein Integrin-β1， p130Cas and α-SMA. Results With 5-days-long amplitude cyclic stretch， cell proliferation and expression of p130Cas， 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 0.1Hz and 10% tensile amplitude 5d can make NSFB exhibit some biochemical characteristics of HSFB， and the mechanical tension can obviously induce the transformation of NSFB to HSFB.
, 百拇医药
Key words： hypertrophic scar； mechanical stress； fibroblasts； cell transformation
皮膚损伤后，创面周围的正常皮肤成纤维细胞(Normal skin fibroblasts， NSFB)增殖迁移，分泌转化生长因子-β(TGF-β)使NSFB向增生性瘢痕成纤维细胞(Hypertrophic scar fibroblasts， HSFB)转化，合成以Ⅰ/Ⅲ型胶原蛋白为主的细胞外基质修复组织，并分泌α-平滑肌肌动蛋白(α-SMA)使组织内纤维收缩，导致增生性瘢痕形成。临床研究发现，切口张力与增生性瘢痕形成呈正相关，表明机械张力是导致增生性瘢痕形成的重要因素[1-2]。实验研究发现，加载机械牵张力后的成纤维细胞有表现出HSFB某些表型的趋势。但是，由于缺乏相应增生性瘢痕成纤维细胞模型，机械张力诱导增生性瘢痕形成的具体机制尚未明确。前期研究表明，加载10%拉伸幅度的周期性机械张力时，NSFB会产生明显的机械传导效应和生化反应[3]，但加载时间对诱导NSFB向HSFB转化过程的影响尚未明确。本实验拟采用机械牵张力诱导NSFB向HSFB转化，观察不同机械牵张力加载时间对NSFB中各生化因子表达水平的影响，探究机械牵张力诱导NSFB向HSFB转化的最佳加载时间，为增生性瘢痕形成机制研究进程奠定前期必要基础，并提供重要实验支撑。, 百拇医药(朱明宇 李雨珊 王志国)
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