何苑琳,郑兆婉,周春霞,洪鹏志

• 1:广东海洋大学食品科技学院/广东省水产品加工与安全重点实验室/广东省海洋食品工程技术研究中心/广东省现代农业科技创新中心/广东省海洋生物制品工程实验室
• 2:广东海洋大学化学与环境学院
• 3:海洋食品精深加工关键技术省部共建协同创新中心大连工业大学

摘要(Abstract)：

【目的】探究从湛江等鞭金藻（Isochrysis zhanjiangensis）中提取的一种五肽AYAPE对UVB照射引起的人永生化角质形成细胞（HaCaT）的抗光老化活性的影响。【方法】CCK-8法测定细胞活力，酶联免疫吸附法ELISA试剂盒测定MMP-1蛋白和Ⅰ型前胶原的分泌变化，免疫组化染色测定p53蛋白的核内表达情况，蛋白质免疫印迹法检测NF-κB信号转导通路中p65蛋白磷酸化的表达及Caspase-3和Caspase-9蛋白的激活程度。【结果】低、中、高浓度下的多肽AYAPE对HaCaT细胞均无毒害作用，且可提高被UVB辐射所损伤的细胞活力；与空白组相比，对照组在UVB的辐射下明显NF-κB信号转导通路被激活，引起细胞凋亡；与对照组相比，样品组中NF-κB信号通路蛋白p65、MMP-1、Cleaved Caspase-3和Cleaved Caspase-9的表达明显降低，而Ⅰ型前胶原的表达明显增加（P <0.05）。【结论】多肽AYAPE通过有效抑制NF-κB信号通路激活、MMP-1分泌、胶原蛋白降解，并显著抑制HaCaT细胞凋亡。

关键词(KeyWords)： HaCaT;抗光老化;UVB;Caspase-3;Caspase-9

Abstract:

Keywords:

基金项目(Foundation): 广东省现代农业产业技术体系创新团队建设项目（2021KJ150）;; 南方海洋科学与工程重点实验室（湛江）资助项目（ZJW-2019-07）;; 湛江市海洋经济创新发展示范市建设项目（XM-202008-01B1）

作者(Author): 何苑琳,郑兆婉,周春霞,洪鹏志

参考文献(References)：

• [1] HABIB M A, SALEM S A M, HAKIM S A, et al.Comparative immunohistochemical assessment of cutaneous cyclooxygenase-2 enzyme expression in chronological aging and photoaging[J]. Photodermatology, Photoimmunology&Photomedicine, 2014, 30(1):43-51.
• [2] SACHS D L, VARANI J, CHUBB H, et al. Atrophic and hypertrophic photoaging:Clinical, histologic, and molecular features of 2 distinct phenotypes of photoaged skin[J].Journal of the American Academy of Dermatology, 2019,81(2):480-488.
• [3] WEN W J, CHEN J W, DING L G, et al. Astragaloside exerts anti-photoaging effects in UVB-induced premature senescence of rat dermal fibroblasts through enhanced autophagy[J]. Archives of Biochemistry and Biophysics,2018, 657:31-40.
• [4] ASSEFA Z, VAN LAETHEM A, GARMYN M, et al.Ultraviolet radiation-induced apoptosis in keratinocytes:on the role of cytosolic factors[J]. Biochimica et Biophysica Acta(BBA)-Reviews on Cancer, 2005, 1755(2):90-106.
• [5] NAZIR L A, TANVEER M A, UMAR S A, et al. Inhibition of ultraviolet-B radiation induced photodamage by trigonelline through modulation of mitogen activating protein kinases and nuclear factor-κB signaling axis in skin[J]. Photochemistry and Photobiology, 2021, 97(4):785-794.
• [6]詹静婷,黄永梅,雷金丽,等.湛江等鞭金藻培养基和培养模式优化的研究[J].基因组学与应用生物学, 2019, 38(3):1248-1255.
• [7] CHEN M F, ZHANG Y Y, MEI D H, et al. Antioxidant peptide purified from enzymatic hydrolysates of Isochrysis zhanjiangensis and its protective effect against ethanol induced oxidative stress of HepG2 cells[J]. Biotechnology and Bioprocess Engineering, 2019, 24(2):308-317.
• [8] YU S J, HU H, ZHENG H, et al. Effect of different phosphorus concentrations on biodiesel production from Isochrysis zhangjiangensis under nitrogen sufficiency or deprivation condition[J]. Applied Microbiology and Biotechnology,2019, 103(12):5051-5059.
• [9] LDONG X L, XUE S, ZHANG J L, et al. The production of light olefins by catalytic cracking of the microalga Isochrysis zhanjiangensis over a modified ZSM-5 catalyst[J]. Chinese Journal of Catalysis, 2014, 35(5):684-691.
• [10] LI Y L, SUN H, WU T, et al. Storage carbon metabolism of Isochrysis zhangjiangensis under different light intensities and its application for co-production of fucoxanthin and stearidonic acid[J]. Bioresource Technology, 2019, 282:94-102.
• [11] CHEN J L, TAN L, LI C Y, et al. Mechanism analysis of a novel angiotensin-I-converting enzyme inhibitory peptide from Isochrysis zhanjiangensis microalgae for suppressing vascular injury in human umbilical vein endothelial cells[J]. Journal of Agricultural and Food Chemistry, 2020, 68(15):4411-4423.
• [12] QIN Z, BALIMUNKWE R M, QUAN T. Age-related reduction of dermal fibroblast size upregulates multiple matrix metalloproteinases as observed in aged human skin in vivo[J]. British Journal of Dermatology, 2017, 177(5):1337-1348.
• [13] ARGYROPOULOS A J, ROBICHAUD P, BALIMUNKWE R M, et al. Alterations of dermal connective tissue collagen in diabetes:molecular basis of aged-appearing skin[J].PLoS One, 2016, 11(4):e0153806.
• [14] FISHER G J, KANG S, VARANI J, et al. Mechanisms of photoaging and chronological skin aging[J]. Archives of Dermatology, 2002, 138(11):1462-1470.
• [15] CHRISTIAN F, SMITH E L, CARMODY R J. The regulation of NF-κB subunits by phosphorylation[J].Cells, 2016, 5(1):12.
• [16] TANAKA K, ASAMITSU K, URANISHI H, et al. Protecting skin photoaging by NF-kappaB inhibitor[J]. Current Drug Metabolism, 2010, 11(5):431-435.
• [17] KIM M J, WOO S W, KIM M S, et al. Anti-photoaging effect of aaptamine in UVB-irradiated human dermal fibroblasts and epidermal keratinocytes[J]. Journal of Asian Natural Products Research, 2014, 16(12):1139-1147.
• [18] WANG L, OH J Y, KIM Y S, et al. Anti-photoaging and anti-melanogenesis effects of fucoidan isolated from Hizikia fusiforme and its underlying mechanisms[J].Marine Drugs, 2020, 18(8):427.
• [19] AYALA M, STRID H, JACOBSSON U, et al. p53expression and apoptosis in the lens after ultraviolet radiation exposure[J]. Investigative Ophthalmology&Visual Science, 2007, 48(9):4187-4191.
• [20] FUJITA E, EGASHIRA J, URASE K, et al. Caspase-9processing by caspase-3 via a feedback amplification loop in vivo[J]. Cell Death&Differentiation, 2001, 8(4):335-344.
• [21] LI P, NIJHAWAN D, BUDIHARDJO I, et al. Cytochrome c and dATP-dependent formation of apaf-1/caspase-9complex initiates an apoptotic protease cascade[J]. Cell,1997, 91(4):479-489.
• [22] CHEN T J, HOU H, FAN Y, et al. Protective effect of gelatin peptides from Pacific cod skin against photoaging by inhibiting the expression of MMPs via MAPK signaling pathway[J]. Journal of Photochemistry and Photobiology B:Biology, 2016, 165:34-41.
• [23] XU D F, WANG W Q, LIAO J M, et al. Walnut protein hydrolysates, rich with peptide fragments of WSREEQEREE and ADIYTEEAGR ameliorate UV-induced photoaging through inhibition of the NF-κB/MMP-1 signaling pathway in female rats[J]. Food&Function, 2020, 11(12):10601-10616.