重组人源Ⅲ型胶原蛋白对豚鼠紫外线辐射损伤皮肤的修复效果探究

doi:10.19586/j.2095-2341.2023.0130

生物技术进展 ›› 2024, Vol. 14 ›› Issue (1): 48-54.DOI: 10.19586/j.2095-2341.2023.0130

• 生物制品研发与技术专题 • 上一篇下一篇

重组人源Ⅲ型胶原蛋白对豚鼠紫外线辐射损伤皮肤的修复效果探究

刁立琴¹(), 李华², 施麟¹, 于月欣², 谢媛¹, 王亚如¹, 徐兰举²()

^1.河北省药品医疗器械检验研究院，石家庄 050227
^2.河北纳科生物科技有限公司，石家庄 050000

收稿日期:2023-10-17 接受日期:2023-12-11 出版日期:2024-01-25 发布日期:2024-02-05
通讯作者: 徐兰举
作者简介:刁立琴 E-mail: 768624340@qq.com；
基金资助:
河北省药品监督管理局科技计划项目(2022ZC1021)

Study on the Repairing Effect of Recombinant Human Type Ⅲ Collagen on Guinea Pig Skin Damaged by Ultraviolet Radiation

Liqin DIAO¹(), Hua LI², Lin SHI¹, Yuexin YU², Yuan XIE¹, Yaru WANG¹, Lanju XU²()

^1.Hebei Institute for Drug and Medical Device Control，Shijiazhuang 050227，China
^2.Hebei NACOL Biotechnology Co. ，Ltd. ，Shijiazhuang 050000，China

Received:2023-10-17 Accepted:2023-12-11 Online:2024-01-25 Published:2024-02-05
Contact: Lanju XU

摘要/Abstract

摘要：

由于大气臭氧层逐渐被破坏，紫外线辐射量大大增加，导致皮肤中的核酸、蛋白质等大分子物质发生化学反应，引起皮肤红斑、色素沉积、晒伤、光老化甚至癌变的发生。以重组人源Ⅲ型胶原蛋白（recombinant human type Ⅲ collagen，rhCol Ⅲ）为研究对象，探究其对成纤维细胞（L929）的细胞毒性及对豚鼠紫外线辐射光毒性损伤皮肤的修复作用。结果发现，在体外细胞实验中rhCol Ⅲ对L929细胞无潜在的细胞毒性；体内动物实验中rhCol Ⅲ对豚鼠紫外线辐射光毒性损伤皮肤具有明显的修复作用，其可以减少豚鼠皮肤损伤后的皮肤皱纹，改善损伤皮肤组织的表皮增生和真皮层炎症反应，降低波形蛋白（Vimentin）和黑色素瘤单克隆抗体45（human melanoma black 45，HMB45）在损伤皮肤组织中的表达。结果表明，rhCol Ⅲ的安全性良好，在豚鼠紫外线辐射损伤皮肤的修复治疗中，可以明显改善紫外辐射豚鼠皮肤的表皮层增厚和角质化程度，降低真皮层处的炎症反应，有效调节成纤维细胞纤维化程度与黑色素瘤的发病风险。结果表明,rhCol Ⅲ对紫外线辐射造成的光损伤皮肤具有明显的治疗效果。

关键词: 重组人源Ⅲ型胶原蛋白, 紫外辐射, 光毒损伤, 皮肤修复效果

Abstract:

Due to the gradual destruction of the ozone layer， the amount of ultraviolet radiation is greatly increased， which would lead to chemical reactions in the skin of nucleic acids， proteins and other large molecules， resulting in skin erythema， pigmentation， sunburn， photoaging and even cancer. Taking recombinant human type Ⅲ collagen （rhCol Ⅲ） as the research object， the cytotoxicity of recombinant human type Ⅲ collagen on fibroblasts （L929） and its repairing effect on skin damaged by ultraviolet radiation in guinea pigs were investigated. The results showed that rhCol Ⅲ had no potential cytotoxicity to L929 cells in vitro. In vivo animal experiments have confirmed that rhCol Ⅲ had obvious repairing effect on the skin damaged by ultraviolet radiation phototoxicity in guinea pigs， which could reduce the skin wrinkles after skin injury， improved the epidermal hyperplasia of the injured skin tissue and the inflammation of the dermis. The expression of Vimentin and human melanoma monoclonal antibody 45 （HMB45） in injured skin were decreased. The above results indicated that rhCol Ⅲ had good safety in the repair treatment of the skin damaged by ultraviolet radiation in guinea pigs， rhCol Ⅲ could significantly improve the degree of epidermal layer thickening and keratinization of the skin of guinea pigs exposed to ultraviolet radiation， reduced the inflammatory response in the dermis and effectively regulate the degree of fibroblast fibrosisand the risk of melanoma. Results showed that， rhCol Ⅲ raw material has obvious therapeutic effect on skin damage caused by ultraviolet radiation.

Key words: recombinant human type Ⅲ collagen, ultraviolet radiation, phototoxic damage, skin repair effect

中图分类号:

Q291

刁立琴, 李华, 施麟, 于月欣, 谢媛, 王亚如, 徐兰举. 重组人源Ⅲ型胶原蛋白对豚鼠紫外线辐射损伤皮肤的修复效果探究[J]. 生物技术进展, 2024, 14(1): 48-54.

Liqin DIAO, Hua LI, Lin SHI, Yuexin YU, Yuan XIE, Yaru WANG, Lanju XU. Study on the Repairing Effect of Recombinant Human Type Ⅲ Collagen on Guinea Pig Skin Damaged by Ultraviolet Radiation[J]. Current Biotechnology, 2024, 14(1): 48-54.

图/表 6

表1 豚鼠皮肤皱纹情况评分标准

Table 1 Scoring criteria for skin wrinkles in guinea pigs

评分标准	评分
正常皮肤细腻纹理	0
皮肤出现细磨痕	1
出现少量浅皱纹	2
出现较多浅皱纹	3
皮肤粗糙，有深皱纹产生	4
有较多粗糙皱纹产生	5
皱纹粗糙，出现皮肤局部损伤	6

图1 rhCol Ⅲ的细胞毒性分析注：**、***表示与空白对照组相比，其他处理组在P<0.01、0.001水平上具有统计学意义。

Fig. 1 Cytotoxicity analysis of rhCol Ⅲ

表2 豚鼠光损伤修复治疗后的皮肤皱纹情况比较

Table 2 Comparison of skin wrinkles in guinea pigs after light damage repair treatment

组别	皮肤皱纹情况评分	t值	显著性（P）
正常组	0.00±0.00	—	—
模型组	4.60±0.55	18.70*	*
1 mg·mL^-1 rhCol Ⅲ组	2.40±0.55	6.33#	#
3 mg·mL^-1 rhCol Ⅲ组	1.60±0.55	8.62#	#
5 mg·mL^-1 rhCol Ⅲ组	0.80±0.45	11.96#	#
市售组	1.40±0.55	9.20#	#

图2 各实验组HE染色分析

Fig. 2 HE staining analysis of each experimental group

图3 各实验组Vimentin免疫组化分析

Fig. 3 Immunohistochemical analysis of Vimentin in each experimental group

图4 各实验组HMB45免疫组化分析

Fig. 4 Immunohistochemical analysis of HMB45 in each experimental group

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重组人源Ⅲ型胶原蛋白对豚鼠紫外线辐射损伤皮肤的修复效果探究

Study on the Repairing Effect of Recombinant Human Type Ⅲ Collagen on Guinea Pig Skin Damaged by Ultraviolet Radiation

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