皮肤光老化小鼠模型的构建及效果评估

doi:10.12300/j.issn.1674-5817.2020.191

摘要/Abstract

摘要： 目的建立中波紫外线诱导小鼠皮肤光老化动物模型,为皮肤光老化的研究提供参考。方法 30只雄性KM小鼠分为对照组和模型组,对照组接受正常光照,模型组使用D-半乳糖皮下注射联合中波紫外线每日照射40 min,每次辐射量为120 mJ/cm²,持续40 d。通过小鼠背部脱毛皮肤表面肉眼观察,皮肤组织HE染色和Masson染色病理学观察,皮肤组织匀浆中谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-PX）、超氧化物歧化酶（superoxide dismutase,SOD）、羟脯氨酸（hydroxyproline,HYP）及丙二醛（malondialdehyde,MDA）含量测定,以及采用蛋白质印迹法检测小鼠背部脱毛皮肤中基质金属蛋白酶1（matrix metalloproteinase-1,MMP-1）、c-fos和c-jun的表达水平,进行建模效果评估。结果与对照组相比,模型组小鼠的皮肤颜色加深、松弛、干燥、粗糙,且褶皱加深;组织病理染色观察发现,模型组的表皮增厚,真皮层纤维减少、断裂、排列杂乱。模型组的皮肤中GSH-PX和SOD 活性降低,HYP含量降低,MDA含量增加,而且MMP-1、c-fos和c-jun表达水平明显升高,差异均有统计学意义（均P<0.01）。结论皮下注射D-半乳糖联合中波紫外线照射后,小鼠产生的皮肤损伤与光老化的皮肤损伤一致;该模型可作为一种有效的皮肤光老化小鼠模型。

关键词: 皮肤光老化, 动物模型, 中波紫外线, D-半乳糖, 小鼠

Abstract: Objective To establish a mouse model of skin photoaging induced by ultraviolet B (UVB) radiation, and to provide a reference for the study of skin photoaging. Methods Thirty male KM mice were divided into a control group and a model group. The control group received normal light, while the model group was subcutaneously injected with D-galactose combined with narrow-band UVB daily irradiation for 40 min at a dose of 120 mJ/cm2 for 40 d. The skin appearance of mice with fur on the back being removed was observed. Skin tissue pathology was observed after HE staining and Masson staining. Biochemical indexes of skin tissue homogenate including glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), hydroxyproline (HYP), and malondialdehyde (MDA) were measured by enzymatic analysis. The expressions of matrix metalloproteinase-1 (MMP-1), c-fos, and c-jun were determined by Western blotting. Results The skin of the model group was darker, looser, drier, more rough, and deeper than that of the control group. Pathological observation revealed that the epidermis thickened and the dermis fibers were reduced, broken, and disorderly arranged in the model group. In the model group, the activities of GSH-PX and SOD decreased, the content of MDA increased, the content of HYP decreased, and the expression levels of MMP-1, c-fos and c-jun of the model group increased (all P<0.01). Conclusion The skin damage of mice induced by subcutaneous injection of D-galactose combined with UVB irradiation is consistent with photoaging, so it is an effective mouse model of skin photoaging.

Key words: Skin photoaging, Animal model, Ultraviolet radiation B, D-galactose, Mice

中图分类号:

R-332
Q95-33

孔悦, 郭砚. 皮肤光老化小鼠模型的构建及效果评估[J]. 实验动物与比较医学, 2021, 41(2): 116-121.

KONG Yue, GUO Yan. Construction and Evaluation of Skin Photoaging Mouse Model[J]. Laboratory Animal and Comparative Medicine, 2021, 41(2): 116-121.

图/表 4

参考文献

[1] GARRE A, NARDA M, VALDERAS-MARTINEZ P, et al.Antiaging effects of a novel facial serum containing L-Ascorbic acid, proteoglycans, and proteoglycan-stimulating tripeptide: ex vivo skin explant studies and in vivo clinical studies in women[J]. Clin Cosmet Investig Dermatol, 2018, 11:253-263. DOI:10.2147/CCID.S161352.
[2] 吴斯敏, 杨慧龄. 紫外线引起皮肤光老化机制及防治的研究进展[J]. 医学综述, 2018, 24(2):341-346. DOI:10.3969/j.issn.1006-2084.2018.02.025.
[3] OLIVEIRA M M, RATTI B A, DAR R G, et al.Dihydrocaffeic acid prevents UVB-induced oxidative stress leading to the inhibition of apoptosis and MMP-1 expression via p38 signaling pathway[J]. Oxid Med Cell Longev, 2019, 2019:2419096. DOI:10.1155/2019/2419096.
[4] SKLAR L R, ALMUTAWA F, LIM H W, et al.Effects of ultraviolet radiation, visible light, and infrared radiation on erythema and pigmentation: a review[J]. Photochem Photobiol Sci, 2013, 12(1):54-64. DOI:10.1039/c2pp25152c.
[5] 龚婕, 马良娟. 皮肤光老化和抗氧化剂研究进展[J]. 中国麻风皮肤病杂志, 2019, 35(11):692-695.
[6] 龚国清, 徐黻本. 小鼠衰老模型研究[J]. 中国药科大学学报, 1991, 22(2):101-103.
[7] AZMAN K F, ZAKARIA R.D-Galactose-induced accelerated aging model: an overview[J]. Biogerontology, 2019, 20(6):763-782. DOI:10.1007/s10522-019-09837-y.
[8] FOLCH J, BUSQUETS O, ETTCHETO M, et al.Experimental models for aging and their potential for novel drug discovery[J]. Curr Neuropharmacol, 2018, 16(10):1466-1483. DOI:10.2174/1570159x15666170707155345.
[9] LIM J Y, KIM O K, LEE J, et al.Protective effect of the standardized green tea seed extract on UVB-induced skin photoaging in hairless mice[J]. Nutr Res Pract, 2014, 8(4):398-403. DOI:10.4162/nrp.2014.8.4.398.
[10] 郭鲁义, 李春雨, 张宁, 等. 实用光老化动物模型建立方法的探讨[J]. 中国美容医学, 2008, 17(2):235-237. DOI:10.3969/j.issn.1008-6455.2008.02.031.
[11] 张璃, 梁虹. 小白鼠皮肤光老化模型的建立[J]. 广东医学, 2005, 26(12):1642-1643. DOI:10.13820/j.cnki.gdyx.2005.12.026.
[12] 杨汝斌, 万屏, 刘玲, 等. SD大鼠光老化模型中氧化性损伤及灯盏花素的保护作用[J]. 中国老年学杂志, 2012, 32(6):1180-1182. DOI:10.3969/j.issn.1005-9202.2012.06.031.
[13] 刘媛, 李福民, 廖金凤, 等. 皮肤光老化机制研究进展[J]. 临床皮肤科杂志, 2016, 45(6):479-481. DOI:10.16761/j.cnki.1000-4963.2016.06.029.
[14] KIM J, LEE C W, KIM E K, et al.Inhibition effect of Gynura procumbens extract on UV-B-induced matrix-metalloproteinase expression in human dermal fibroblasts[J]. J Ethnopharmacol, 2011, 137(1):427-433. DOI:10.1016/j.jep.2011.04.072.
[15] MUTHUSAMY V, PIVA T J.The UV response of the skin: a review of the MAPK, NFkappaB and TNFalpha signal transduction pathways[J]. Arch Dermatol Res, 2010, 302(1):5-17. DOI:10.1007/s00403-009-0994-y.
[16] VICENTINI F T, HE T Y, SHAO Y, et al.Quercetin inhibits UV irradiation-induced inflammatory cytokine production in primary human keratinocytes by suppressing NF-κB pathway[J]. J Dermatol Sci, 2011, 61(3):162-168. DOI:10.1016/j.jdermsci.2011.01.002.
[17] YU W, YING H H, TONG F D, et al.Protective effect of the silkworm protein 30Kc6 on human vascular endothelial cells damaged by oxidized low density lipoprotein (Ox-LDL)[J]. PLoS One, 2013, 8(6): e68746. DOI:10.1371/journal.pone.0068746.
[18] TAKAHASHI Y, FUKUSHIMA Y, KONDO K, et al.Facial skin photo-aging and development of hyperpigmented spots from children to middle-aged Japanese woman[J]. Skin Res Technol, 2017, 23(4):613-618. DOI:10.1111/srt.12380.
[19] 顾振华, 汪俊军. 基质金属蛋白酶-1的研究进展[J]. 医学研究生学报, 2006, 19(11):1028-1031. DOI:10.16571/j.cnki.1008-8199.2006.11.022.
[20] FISHER G J, TALWAR H S, LIN J, et al.Molecular mechanisms of photoaging in human skin in vivo and their prevention by all-trans retinoic acid[J]. Photochem Photobiol, 1999, 69(2):154-157. DOI:10.1562/0031-8655(1999)0690154: mmopih>2.3.co;2.
[21] 邓映, 杜宇, 刘萍, 等. 氧化应激在皮肤光老化中的作用[J]. 中国医疗美容, 2020, 10(8):117-122. DOI:10.19593/j.issn.2095-0721.2020.08.030.
[22] 林芳. 金属蛋白酶在皮肤光老化信号通路中的研究进展[J]. 临床与病理杂志, 2018, 38(11):2502-2507. DOI:10.3978/j.issn.2095-6959.2018.11.033.