激光生物学报, 2018, 27 (5): 399, 网络出版: 2018-11-25  

重组人源胶原蛋白对激光损伤小鼠皮肤修复作用及机制研究

Repair Effect and Mechanism of Recombinant Human Collagen on Laser Induced Damage of Mice Skin
作者单位
1 中国中医科学院医学实验中心, 北京 100700
2 中医药防治重大疾病基础研究北京市重点实验室, 北京 100700
3 天津伊瑞雅生物科技有限公司, 天津 300399
4 天津中新药业集团股份有限公司中新制药厂, 天津 300380
摘要
本研究旨在观察重组人源胶原蛋白(recombinant human collagen,r-hc)对小鼠皮肤激光损伤的修复作用, 初步探索其作用机制。应用458~514 nm激光照射小鼠背部皮肤制作皮肤损伤模型, 将r-hc外涂于创伤皮肤, 剂量为 8 mg/mL(生理盐水配制), 每天涂抹1次, 连续给药14 d。分别于给药后1、4、7、14 d用双光子显微镜收集二次谐波(second harmonic generation,SHG)信号检测伤口真皮中的胶原纤维, 并进行常规 HE 染色, 观察伤口局部的病理学改变。体外试验检测r-hc对人皮肤角质形成细胞和成纤维细胞增殖活力的影响, 计算细胞存活率。在小鼠模型上显示, 与对照组相比, r-hc可明显加速伤口的愈合, 缩短伤口愈合时间; SHG显示 r-hc能够促进创伤局部胶原的产生; 体外试验也显示它有促进人皮肤成纤维细胞和角质形成细胞增殖的能力。由此可见, r-hc(8 mg/mL)对小鼠激光损伤皮肤有修复作用, 可能是通过促进表皮角质细胞和真皮成纤维细胞的增殖, 促进胶原的沉积而发挥修复作用的。
Abstract
This study was to observe the repair effect of recombinant human collagen(r-hc)on laser induced damage of mice skin and to explore preliminarily its mechanism.Dorsal skin of mice was irradiated with 458~514 nm laser, r-hc dissolved in physiological saline solution(8 mg/mL)was used to treat dorsal skin wounds of mice, once per day for 14 days. The collagen fibers in the wound dermis were observed by using the backwards second harmonic generation(SHG)signals which were collected by two photon microscopy(1,4,7,14 d)after the administration. The pathological changes of the wound were observed by routine HE staining. The effects of r-hc on proliferation of epidermic cells and fibroblasts were studied in vitro, and cell viability was calculated.The results indicated that compared with the control group, r-hc significantly accelerated wound healing and shortened wound healing time in mice. SHG imaging showed that r-hc could promote the production of collagen in wound, and in vitro experiments r-hc showed markedly proliferation-promoting activity for human skin fibroblasts and keratinocytes. It can be concluded that r-hc(8 mg/mL)has a repairing effect on laser-damaged mouse skin, which may be achieved by stimulating the proliferation of epidermal keratinocytes and dermal fibroblasts and promoting collagen deposition.

马淑骅, 孙娅楠, 杨伟峰, 孙桂荣, 李松林, 蒋盈宝, 王岩, 王毅. 重组人源胶原蛋白对激光损伤小鼠皮肤修复作用及机制研究[J]. 激光生物学报, 2018, 27(5): 399. MA Shuhua, SUN Yanan, YANG Weifeng, SUN Guirong, LI Songlin, JIANG Yingbao, WANG Yan, WANG Yi. Repair Effect and Mechanism of Recombinant Human Collagen on Laser Induced Damage of Mice Skin[J]. Acta Laser Biology Sinica, 2018, 27(5): 399.

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