重组人源化胶原蛋白基于体外水平的护肤功效研究

doi:10.3969/j.issn.2097-2806.2024.09.003

摘要/Abstract

摘要：

基于体外水平探究重组人源化胶原蛋白的护肤功效。首先通过不同温度对重组人源化胶原蛋白进行处理，分析其在不同温度条件下的稳定性，随后通过检测其对弹性蛋白酶的抑制率、胶原蛋白酶的抑制率、人成纤维细胞Ⅰ型胶原蛋白含量、体外角质形成细胞活性氧（ROS）抑制作用及对角质形成细胞透明质酸（HA）、丝聚蛋白（FLG）和转谷氨酰胺酶1（TGM1）表达量的影响，评价重组人源化胶原蛋白的体外护肤功效。结果显示，重组人源化胶原蛋白在70 ℃以内能保持其稳定性良好。在护肤功效方面，重组人源化胶原蛋白能够抑制弹性蛋白酶和胶原蛋白酶的活性，促进人成纤维细胞Ⅰ型胶原蛋白含量上升。同时，其对体外角质形成细胞ROS具有显著的抑制作用，并可以增加角质形成细胞HA、FLG和TGM1的表达量。总之，重组人源化胶原蛋白具有良好的体外护肤功效，本研究证明其具有潜在的紧致、抗皱、保湿及修护作用，是一种理想的化妆品原料。

关键词: 重组人源化胶原蛋白, 稳定性, 人成纤维细胞, 体外角质形成细胞, 护肤功效

Abstract:

Studying the skin care efficacy of recombinant humanized collagen based on in vitro level. The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures, then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase, the inhibition rate of collagenase, the protein content of type I collagen in human fibroblasts, the inhibition of reactive oxygen species （ROS） with human keratinocytes, and the effects of the recombinant humanized collagen on the expression of hyaluronic acid （HA）, filaggrin （FLG） and transglutaminase 1 （TGM1） in keratinocytes. The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70 ℃. With regard to its skincare efficacy, recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts. It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA, FLG, and TGM1 in keratinocytes. In short, the recombinant humanized collagen exhibited a favourable skin care effect in vitro level. This study proved that it has potential firming, anti-wrinkle, moisturizing, and repairing efficacy, and is a valuable cosmetic raw material.

Key words: recombinant humanized collagen, stability, human fibroblast cell, in vitro keratinocytes, skin care efficacy

中图分类号:

TQ658

王建, 范毓慧, 李丹凤, 程宁文, 李玲, 于玉凤. 重组人源化胶原蛋白基于体外水平的护肤功效研究[J]. 日用化学工业（中英文）, 2024, 54(9): 1030-1038.

Jian Wang, Yuhui Fan, Danfeng Li, Ningwen Cheng, Ling Li, Yufeng Yu. Skin care efficacy study of recombinant humanized collagen based on in vitro level[J]. China Surfactant Detergent & Cosmetics, 2024, 54(9): 1030-1038.

图/表 10

参考文献 23

[1]	Li Jie, Wang Mingchao, Qiao Yingyun, et al. Extraction and characterization of type Ⅰ collagen from skin of tilapia (Oreochromis niloticus) and its potential application in biomedical scaffold material for tissue engineering[J]. Process Biochemistry, 2018, 74: 156-163.
[2]	Fan Ting, Zhao Jianfeng, Chang Yejun, et al. Effect of recombinant humanized type Ⅲ collagen on expression of skin functional related genes[J]. China Surfactant Detergent & Cosmetics, 2022, 52 (12): 1326-1332.
[3]	Zhou Yuhan, Yuan Rongsui, Jiang Shoujun, et al. Biomedical application and development prospect of recombinant collagen[J]. Shanxi Chemical Industry, 2023, 43 (12): 29-31.
[4]	Chen Zhuoyue, Fan Daidi, Shang Lijun. Exploring the potential of the recombinant human collagens for biomedical and clinical applications: a short review[J]. Biom-edical Materials, 2021, 16 (1): 012001.
[5]	Ma Shuhua, Sun Yanan, Yang Weifeng, et al. Repair effect and mechanism of recombinant human collagen on laser induced damage of mice skin[J]. Acta Laser Biology Sinica, 2018, 27 (5): 399-406.
[6]	Yang Jianqiu, Gao Yihong, Zhu Hongliu. Therapeutic effect and mechanism of recombinant human type Ⅲ collagen functional gel on skin wound healing[J]. Shandong Medical Journal, 2021, 61 (34): 80-83.
[7]	Wang Jing, Qiu He, Xu Yang, et al. The biological effect of recombinant humanized collagen on damaged skin induced by UV-photoaging: an in vivo study[J]. Bioactive Materials, 2022, 11: 154-165. doi: 10.1016/j.bioactmat.2021.10.004 pmid: 34938920
[8]	Liu Hongye, Guo Shuping, Bai Li. Clinical observation of medical recombinant human collagen functional dressing for wound repair after carbon dioxide laser surgery[J]. Chinese Remedies & Clinics. 2015, 15 (10): 1458-1460.
[9]	Mao Zhengwei, Gao Changyou, Wang Dengyou, et al. Controlling biostability of collagen films for fibroblast cytocompatibility[J]. Journal of Bioactive and Compatible Polymers, 2004, 19 (5): 353-365.
[10]	Mu Changdao. A collagen facial mask and its preparation method: CN102138870A[P]. 2011-08-03.
[11]	Zhang Hui. Expression and purification of recombinant human collagen-like and its application in cosmetics[D]. Guangzhou: Jinan University, 2017.
[12]	He Xingxing, Zhao Yue, Lu Chaogang, et al. Study on the efficacy and evaluation of collagen, elastin and fibronectin in cosmetic[J]. Fine and Specialty Chemical, 2023, 31 (10): 39-43.
[13]	Wang Yulin. Research progress on the application of collagen in cosmetics[J]. The Science and Technology of Gelatin, 2012, 32 (1): 8-12.
[14]	Liang Meizhen. The application of protein in cosmetics[J]. Guangdong Chemical Industry, 2009, 36 (12): 104-105.
[15]	Lu Huanhuan, Zhou Xuye, Zhang Zhiwu. Study on effect of collagen type Ⅰ on tissue regeneration and anti-inflammation in zebrafish model[J]. Chinese Journal of Modern Applied Pharmacy, 2022, 39 (9): 1182-1186.
[16]	Nguyen T T, Happillon T, Feru J, et al. Raman comparison of skin dermis of different ages: focus on spectral markers of collagen hydration[J]. Journal of Raman Spectroscopy, 2013, 44 (9): 1230-1237.
[17]	Bodin J, Adrien A, Bodet P E, et al. Ulva intestinalis protein extracts promote in vitro collagen and hyaluronic acid production by human dermal fibroblasts[J]. Molecules, 2020, 25 (9): 2091.
[18]	Gu Yanpei, Han Jianxin, Jiang Chunpeng, et al. Biomarkers, oxidative stress and autophagy in skin aging[J]. Ageing Research Reviews, 2020, 59 (19): 101036.
[19]	Kanaki T, Makrantonaki E, Zouboulis C C. Biomarkers of skin aging[J]. Reviews in Endocrine & Metabolic Disorders, 2016, 17 (3): 433-442.
[20]	Tanha A, Rabiee M, Rostami A, et al. A green-based approach for noninvasive skin rejuvenation: potential application of hyaluronic acid[J]. Environmental Research, 2023, 234: 116467.
[21]	Li Wenhwa, Wong Hengkuan, Serrano J, et al. Topical stabilized retinol treatment induces the expression of HAS genes and HA production in human skin in vitro and in vivo[J]. Archives of Dermatological Research, 2017, 309 (4): 275-283. doi: 10.1007/s00403-017-1723-6 pmid: 28247017
[22]	Lai Q W S, Guo M S S, Wu K Q, et al. Edible bird’s nest, an asian health food supplement, possesses moisturizing effect by regulating expression of filaggrin in skin keratinocyte[J]. Frontiers in Pharmacology, 2021, 12: 685982.
[23]	Wolf C, Qian Y, Brooke M A, et al. ADAM17/EGFR axis promotes transglutaminase-dependent skin barrier formation through phosholipase Cγ1 and protein kinase C pathways[J]. Scientific Reports, 2016, 6: 39780.

	φ（RHC）/%
	100	80	60	40	20	10	1	0.1
Relative cell viability/%	2.50	3.30	20.92	47.35	81.88	95.50	98.20	101.40
SD	0.00	0.00	0.01	0.02	0.01	0.01	0.00	0.01

	φ（RHC）/%
	100	80	60	40	20	10	1	0.1
Relative cell viability/%	0.65	4.45	26.71	59.93	88.82	98.05	103.91	103.80
SD	0.00	0.00	0.00	0.01	0.01	0.01	0.00	0.01

Groups	ρ（Collagen type Ⅰ）/（pg/mL）	Collagen type Ⅰ increase rate/%	P-value
Negative Control	470.04±20.12	—	—
Positive Control	776.95±17.12	65.29	**
φ（RHC） =0.1%	772.24±27.92	64.29	**
φ（RHC） =1%	819.57±29.48	74.36	**
φ（RHC） =10%	876.83±22.00	86.54	**