[不同幅度周期性拉伸对正常皮肤成纤维细胞向增生性瘢痕成纤维细胞转化的影响]

[Effect of cyclic stretch with different amplitudes on the transformation of normal skin fibroblasts into hypertrophic scar fibroblasts].

作者信息

Wang Zhiguo, Kuang Ruixia, Chen Zhenyu, Li Peng, Chen Lu, Zhang Weidong

机构信息

Department of Plastic Surgery, Affiliated Hospital, Qingdao University, Qingdao266003, China. Email:

Department of Plastic Surgery, Affiliated Hospital, Qingdao University, Qingdao266003, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2015 Jan 27;95(4):294-8.

PMID:25877248

Abstract

OBJECTIVE

To explore the role of cyclic stretch with different amplitudes on the transformation of normal skin fibroblasts (NSFB) into hypertrophic scar fibroblasts (HSFB).

METHODS

NSFB and HSFB were cultured in vitro. Cyclic stretch with 10%, 15% and 20% amplitude was applied for 24 h on NSFB of experimental groups (Group N10%,Group N15% and Group N20%). And control groups (groups N0 and H0), NSFB and HSFB were cultured without stretch. Cell Counting Kit-8 method was used to detect the proliferation of fibroblasts. The gene expressions of integrin β1, P130Cas, TGF-β1, COL1A1 and COL3 A1 were detected by real-time polymerase chain reaction (PCR). And the protein expressions of TGF-β1, collagen types I and III were detected by enzyme-linked immunosorbent assay.

RESULTS

Cyclic stretch had stimulatory effect on cell proliferation. The proliferation capacity of group N10% was higher than that of group N20% (0.91 ± 0.09 vs 0.8 ± 0.07, P < 0.05). Cyclic stretch could induce increased levels of gene expression.Gene expression levels of integrin β1, P130Cas, TGF-β1, COL1A1 and COL3 A1 in group N10% was higher than those in group N20% (3.14 ± 0.24, 5.01 ± 0.49, 1.55 ± 0.08, 2.38 ± 0.08, 1.37 ± 0.11 vs 1.78 ± 0.32, 3.07 ± 0.41, 1.18 ± 0.05, 1.97 ± 0.05, 1.26 ± 0.19; all P < 0.05). Cyclic stretch could stimulate NSFB to secrete TGF-β1. The protein level of TGF-β1 in group N10% was higher than that in group N20% ((459 ± 12.96) vs (372 ± 10.49) pg/ml, P < 0.05). The protein levels of collagen types I and III in experiment groups were higher than those in group N0 and lower than those in group H0 (all P < 0.05). The protein level of collagen type I in group N10% was higher than that in group N20% ((12.66 ± 1.16) vs (11.23 ± 0.75) ng/ml, P < 0.05).

CONCLUSIONS

NSFB loaded cyclic stretch with 10% amplitude could exhibit some biological characteristics of HSFB. And cyclic stretch with optimal amplitude might transform NSFB into HSFB.

摘要

目的

探讨不同幅度的周期性拉伸对正常皮肤成纤维细胞（NSFB）向增生性瘢痕成纤维细胞（HSFB）转化的作用。

方法

体外培养NSFB和HSFB。对实验组（N10%组、N15%组和N20%组）的NSFB施加幅度为10%、15%和20%的周期性拉伸24小时。而对照组（N0组和H0组），NSFB和HSFB在无拉伸条件下培养。采用细胞计数试剂盒-8法检测成纤维细胞的增殖情况。通过实时聚合酶链反应（PCR）检测整合素β1、P130Cas、转化生长因子-β1（TGF-β1）、I型胶原蛋白（COL1A1）和III型胶原蛋白（COL3A1）的基因表达。并通过酶联免疫吸附测定法检测TGF-β1、I型和III型胶原蛋白的蛋白表达。

结果

周期性拉伸对细胞增殖有刺激作用。N10%组的增殖能力高于N20%组（0.91±0.09对0.8±0.07，P<0.05）。周期性拉伸可诱导基因表达水平升高。N10%组中整合素β1、P130Cas、TGF-β1、COL1A1和COL3A1的基因表达水平高于N20%组（3.14±0.24、5.01±0.49、1.55±0.08、2.38±0.08、1.37±0.11对1.78±0.32、3.07±0.41、1.18±0.05、1.97±0.05、1.26±0.19；均P<0.05）。周期性拉伸可刺激NSFB分泌TGF-β1。N10%组中TGF-β1的蛋白水平高于N20%组（（459±12.96）对（372±10.49）pg/ml，P<0.05）。实验组中I型和III型胶原蛋白的蛋白水平高于N0组且低于H0组（均P<0.05）。N10%组中I型胶原蛋白的蛋白水平高于N20%组（（12.66±1.16）对（11.23±0.75）ng/ml，P<0.05）。