丝素蛋白对皮肤光损伤的保护作用

doi:10.3969/j.issn.2097-2806.2024.09.013

摘要/Abstract

摘要：

丝素蛋白是一种从蚕丝中提取的天然高分子纤维蛋白，有很好的抗氧化和减少光损伤的作用。为了研究丝素蛋白对皮肤光损伤的保护作用，以UVB诱导人角质形成细胞（HaCaT）损伤为模型，通过ELISA和荧光检测细胞内活性氧（ROS）和钙离子（Ca²⁺）的水平，采用免疫荧光检测瞬时受体电位阳离子通道亚家族V成员1 （TRPV1）和Claudin-1的表达。此外，还使用蛋白质印迹分析了紧密连接相关蛋白Occludin和Claudin-1的表达。结果表明，120 mJ/cm²的UVB刺激显著降低细胞活力，而100 μg/mL丝素蛋白的存在显著增加细胞活力。UVB刺激导致细胞内活性氧水平升高，激活TRPV1通道，诱导细胞内Ca²⁺水平增加。同时炎症因子白细胞介素1α （IL-1α）（P<0.01）和S100钙结合蛋白A8 （S100A8）（P<0.05）水平也显著升高，引发炎症反应。添加丝素蛋白的HaCaT细胞受到UVB的影响则显著降低（P<0.05）。实验证明，UVB会破坏屏障蛋白，导致屏障蛋白Claudin-1和Occludin的表达下降。丝素蛋白的加入可以降低这种影响，保护细胞间紧密连接。因此，丝素蛋白可能具有降低氧化应激、屏障损伤和炎症发生，保护皮肤免受UVB诱导的光损伤的潜在能力。

关键词: 丝素蛋白, UVB, 活性氧, 紧密连接蛋白, 皮肤屏障

Abstract:

As a natural macromolecular protein extracted from silk, silk fibroin （SF） has been widely used as biological material in recent years due to its antioxidant and photodamage reduction effects. To study the protective effect of silk fibroin against skin photodamage, the immortalized human keratinocyte line （HaCaT） was used as a model for study of UVB injury. Intracellular reactive oxygen species （ROS） and the levels of calcium ions （Ca²⁺） were measured by ELISA and fluorescence microscopy. The expression of transient receptor potential cation channel subfamily V member 1 （TRPV1） and the expression of Claudin-1 were detected by immunofluorescence assay. In addition, western blotting was used to analyze the expression of tight junction proteins occludin and Claudin-1. The results showed that 120 mJ/cm² of UVB stimulation significantly reduced cell viability, while the presence of 100 μg/mL silk fibroin significantly increased cell viability. UVB stimulation could increase the level of intracellular ROS, activate TRPV1 channels, and induce the increase of intracellular Ca²⁺ level. Meanwhile, the levels of inflammatory cytokines interleukin-1α （IL-1α）（P<0.01） and S100A8 （P<0.05） were also significantly increased, which caused inflammation. However, the effect of UVB on HaCaT cells with the addition of silk fibroin was significantly decreased （P<0.05）. In conclusion, UVB could disrupt the structural integrity of barrier proteins, resulting in the decreased expression of the barrier proteins Claudin-1 and occludin. The addition of silk fibroin could reduce such effect and protect the tightly connected structure. Therefore, silk fibroin might have great potential to protect skin from UV-induced inflammation, barrier damage and oxidative stress damage.

Key words: silk fibroin, UVB, ROS, tight-junction protein, skin barrier

中图分类号:

TQ658

李生鹏, 李静, 梁超, 赵冉, 张晓洁, 孙丽丽. 丝素蛋白对皮肤光损伤的保护作用[J]. 日用化学工业（中英文）, 2024, 54(9): 1117-1124.

Shengpeng Li, Jing Li, Chao Liang, Ran Zhao, Xiaojie Zhang, Lili Sun. Protective effect of silk fibroin on UVB-induced skin injury[J]. China Surfactant Detergent & Cosmetics, 2024, 54(9): 1117-1124.

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参考文献 38

[1]	Prost-Squarcioni C, Histologie De La Peauet, Des Follicules Pileux. Histology of skin and hair follicle[J]. Med Sci (Paris), 2006, 22 (2): 131-137.
[2]	Basler K, Bergmann S, Heisig M, et al. The role of tight junctions in skin barrier function and dermal absorption[J]. J Control Release, 2016, 242: 105-118.
[3]	Dang N N, Pang S G. Filaggrin silencing by shRNA directly impairs the skin barrier function of normal human epidermal keratinocytes and then induces an immune response[J]. Braz J Med Biol Res, 2015, 48 (1): 39-45. doi: S0100-879X2015000100039 pmid: 25493381
[4]	Boivin F J, Schmidt-Ott K M. Transcriptional mechanisms coordinating tight junction assembly during epithelial differentiation[J]. Ann N Y Acad Sci, 2017, 1397 (1): 80-99.
[5]	Pummi K, Malminen M. Epidermal tight junctions: ZO-1 and occludin are expressed in mature, developing, and affected skin and in vitro differentiating keratinocytes[J]. J Invest Dermatol, 2001, 117 (5): 1050-1058. pmid: 11710912
[6]	Tu Y, Wang L, Wang X, et al. LncRNA-WAKMAR2 regulates expression of CLDN1 to affect skin barrier through recruiting c-Fos[J]. Contact Dermatitis, 2023, 88 (3): 188-200.
[7]	Prade JdS, Bálsamo E C. Anti-inflammatory effect of Arnica montana in a UVB radiation-induced skin-burn model in mice[J]. Cutan Ocul Toxicol, 2020, 39 (2): 1-23.
[8]	Garmyn M, Young A, Miller S. Mechanisms of and variables affecting UVR photoadaptation in human skin[J]. Photochem Photobiol Sci, 2018, 17 (12): 1932-1940.
[9]	Peharda V, Gruber F, Kastelan M. Occupational skin diseases caused by solar radiation[J]. Collegium Antropologicum, 2007, 15: 87-90.
[10]	Tanaka Y, Uchi H, Furue M. Antioxidant cinnamaldehyde attenuates UVB-induced photoaging[J]. J Dermatol Sci, 2019, 96 (3): 151-158. doi: S0923-1811(19)30340-8 pmid: 31735467
[11]	D’Orazio J, Jarrett S. UV radiation and the skin[J]. Int J Mol Sci, 2013, 14 (6): 12222-12248. doi: 10.3390/ijms140612222 pmid: 23749111
[12]	Matsumura Y, Ananthaswamy H N. Toxic effects of ultraviolet radiation on the skin[J]. Toxicol Appl Pharmacol, 2004, 195 (3): 298-308.
[13]	Schonthaler H B, Guinea-Viniegra J. S100A8-S100A9 protein complex mediates psoriasis by regulating the expression of complement factor C3[J]. Immunity, 2013, 39 (6): 1171-1181. doi: 10.1016/j.immuni.2013.11.011 pmid: 24332034
[14]	Wang S, Song R. S100A8/A9 in Inflammation[J]. Front Immunol, 2018, 11 (9): 1298.
[15]	Draelos Z D. The cosmeceutical realm[J]. Clin Dermatol, 2008, 26 (6): 627-632. doi: 10.1016/j.clindermatol.2007.09.005 pmid: 18940543
[16]	Vepari C, Kaplan D L. Silk as a biomaterial[J]. Prog Polym Sci, 2007, 32 (8/9): 991-1007.
[17]	Kasoju N, Bora U. Silk fibroin in tissue engineering[J]. Adv Healthc Mater, 2012, 1 (4): 393-412.
[18]	Yan S, Zhang Q, Wang J, et al. Silk fibroin/chondroitin sulfate/hyaluronic acid ternary scaffolds for dermal tissue reconstruction[J]. Acta Biomater, 2013, 9 (3): 6771-6782.
[19]	Jeong L, Kim M H, Jung J Y, et al. Effect of silk fibroin nanofibers containing silver sulfadiazine on wound healing[J]. Int J Nanomedicine, 2014, 9: 5277-5287.
[20]	Dash R, Mandal M. Silk sericin protein of tropical tasar silkworm inhibits UVB-induced apoptosis in human skin keratinocytes[J]. Mol Cell Biochem, 2008, 311: 111-119. doi: 10.1007/s11010-008-9702-z pmid: 18214642
[21]	Berardesca E, Ardigo M. Randomized, double-blinded, vehicle-controlled, split-face study to evaluate the effects of topical application of a Gold Silk Sericin/Niacinamide/Signaline complex on biophysical parameters related to skin ageing[J]. Int J Cosmet Sci, 2015, 37: 606-612.
[22]	Barajas-Gamboa J A, Serpa-Guerra A M. Aplicaciones de la sericina: Una proteina globular proveniente de la seda[J]. Ing Compet, 2016, 18: 193-206.
[23]	Chlapanidas T, Faragò S, Lucconi G, et al. Sericins exhibit ROS-scavenging, anti-tyrosinase, anti-elastase, and in vitro immunomodulatory activities[J]. Int J Biol Macromol, 2013, 58: 47-56. doi: 10.1016/j.ijbiomac.2013.03.054 pmid: 23541552
[24]	Padamwar M N, Pawar A P. Silk sericin and its applications: A review[J]. J Sci Ind Res, 2004, 63: 323-329.
[25]	Dash R, Mandal M, Ghosh S K, et al. Silk sericin protein of tropical tasar silkworm inhibits UVB-induced apoptosis in human skin keratinocytes[J]. Mol Cell Biochem, 2008, 311 (1/2): 111-119.
[26]	Kammeyer A, Luiten R M. Oxidation events and skin aging[J]. Ageing Research Reviews, 2015, 21: 16-29. doi: 10.1016/j.arr.2015.01.001 pmid: 25653189
[27]	Choi E H. Aging of the skin barrier[J]. Clin Dermatol, 2019, 37 (4): 336-345. doi: S0738-081X(19)30076-8 pmid: 31345321
[28]	Choi H J, Alam M B, Baek M E, et al. Protection against UVB-induced photoaging by Nypa fruticans via inhibition of MAPK/AP-1/MMP-1 signaling[J]. Oxid Med Cell Longev, 2020, 2020:2905362.
[29]	Manosroi A, Boonpisuttinant K. Free radical scavenging and tyrosinase inhibition activity of oils and sericin extracted from Thai native silkworms (Bombyx mori)[J]. Pharm Biol, 2010, 48 (8): 855-860. doi: 10.3109/13880200903300212 pmid: 20673171
[30]	Choi J H, Lee S. Antioxidation and anti-inflammatory effects of gamma-irradiated silk sericin and fibroin in H₂O₂-induced HaCaT cell[J]. Korean J Physiol Pharmacol, 2023, 27 (1): 105-112.
[31]	Yoo J A, Yu E, Park S H, et al. Blue light irradiation induces human keratinocyte cell damage via transient receptor potential vanilloid 1(TRPV1) regulation[J]. Oxid Med Cell Longev, 2020, 2020: 8871745.
[32]	Kang S M, Han S. A synthetic peptide blocking TRPV1 activation inhibits UV-induced skin responses[J]. J Dermatol Sci, 2017, 88 (1): 126-133. doi: S0923-1811(17)30074-9 pmid: 28551094
[33]	Dinarello C A. Interleukin-1[J]. Ann N Y Acad Sci, 1988, 546: 122-132.
[34]	Zhang Chen, Yang Ya. Effect of Lycium barbarum polysaccharide on expression of inflammatory factors in keratinocytes after UVB radiation[J]. Chinese Medical Review, 2017, 14 (19): 20-23.
[35]	Santamaria-Kisiel L, Rintala-Dempsey A C, Shaw G S. Calcium-dependent and-independent interactions of the S100 protein family[J]. Biochem J, 2006, 396 (2): 201-214. doi: 10.1042/BJ20060195 pmid: 16683912
[36]	Gebhardt C, Németh J. S100A8 and S100A9 in inflammation and cancer[J]. Biochem Pharmacol, 2006, 72 (11): 1622-1631. doi: 10.1016/j.bcp.2006.05.017 pmid: 16846592
[37]	Xu C L, Sun R. Protective effect of glutamine on intestinal injury and bacterial community in rats exposed to hypobaric hypoxia environment[J]. World J Gastroenterol, 2014, 20 (16): 4662-4674.
[38]	Li Y, Liu J, Pongkorpsakol P, et al. Relief effects of icariin on inflammation-induced decrease of tight junctions in intestinal epithelial cells[J]. Front Pharmacol, 2022, 13: 903762.