浙江红山茶籽油酶解产物抗D-gal诱导皮肤成纤维细胞衰老作用及机制研究

doi:10.3969/j.issn.2097-2806.2026.05.008

摘要/Abstract

摘要：

探讨浙江红山茶籽油酶解产物（enzymatic hydrolysis products of Camellia chekiangoleosa Hu seed oil, EHCO）对D-gal诱导人皮肤成纤维细胞（human normal foreskin fibroblasts, HFF）衰老的保护作用及机制研究。使用60 mg/mL D-gal建立HFF细胞衰老模型，D-gal和体积分数0.1%，0.5% EHCO共同处理HFF细胞24 h，检测细胞活力和衰老水平。ELISA试剂盒检测ATP和SOD含量，ROS、JC-1、EDU荧光染色检测细胞线粒体功能和增殖水平，RT-qPCR检测衰老和炎症相关基因的表达，Western blotting检测凋亡和细胞自噬相关蛋白的表达。EHCO处理的HFF细胞内P16、P21、P53、IL-1β、IL-6 mRNA水平以及P53、ERK的磷酸化水平呈剂量依赖性降低（P<0.05），同时BAX与BCL-2蛋白比值减小，自噬相关蛋白（LC3B）表达升高。本研究发现生物脂肪酶切技术对红山茶籽油进行酶解获得的EHCO能够促进细胞增殖，对抗D-gal诱导的HFF细胞衰老。机制上，EHCO通过ERK-P53信号轴和激活细胞自噬改善成纤维细胞功能，来发挥抗皮肤衰老的作用。

关键词: 浙江红山茶籽油酶解产物, 成纤维细胞, 细胞自噬, 衰老

Abstract:

This study investigated the protective effects and underlying mechanisms of the enzymatic hydrolysate from Camellia chekiangoleosa Hu seed oil （EHCO） on D-galactose （D-gal）- induced senescence in human normal foreskin fibroblasts （HFF）. A cellular senescence model was established by treating HFFs with 60 mg/mL D-gal. Cells were co-treated with D-gal and EHCO （0.1% or 0.5%） for 24 hours, and cell viability and senescence levels were subsequently assessed. The content of ATP and the activity of SOD were measured using ELISA kits. Mitochondrial function and proliferative capacity were evaluated via fluorescent staining for ROS, JC-1, and EDU. The mRNA expression levels of senescence-associated and inflammation-related genes were quantified by RT-qPCR. Furthermore, the protein expression of apoptosis-related and autophagy-related markers, along with the phosphorylation status of P53 and ERK, were analyzed by Western blotting. In EHCO-treated HFF, the mRNA levels of P16, P21, P53, IL-1β, and IL-6, as well as the phosphorylation levels of P53 and ERK, significantly reduce in a dose-dependent manner （P<0.05）. Concurrently, the pro-apoptotic BAX/BCL-2 protein ratio decreases, while the expression of the autophagy marker protein LC3B elevates. This study demonstrates that EHCO, obtained through lipase-catalyzed hydrolysis of Camellia chekiangoleosa Hu, promotes cell proliferation and protects against D-gal-induced senescence in HFF. Mechanistically, EHCO exerts anti-skin-aging effects by improving fibroblast function via the suppression of the ERK-P53 signaling axis and the activation of cellular autophagy.

Key words: enzymatic hydrolysate of Camellia chekiangoleosa Hu seed oil, fibroblasts, autophagy, senescence

中图分类号:

TQ658

卢琦, 向文叠, 李广涛, 尹世鹏, 高宏旗, 余果. 浙江红山茶籽油酶解产物抗D-gal诱导皮肤成纤维细胞衰老作用及机制研究[J]. 日用化学工业（中英文）, 2026, 56(5): 614-622.

Qi Lu, Wendie Xiang, Guangtao Li, Shipeng Yin, Hongqi Gao, Guo Yu. Anti-aging effects and mechanism of enzymatic hydrolysate of Camellia chekiangoleosa Hu seed oil against D-galactose-induced senescence in human skin fibroblasts[J]. China Surfactant Detergent & Cosmetics, 2026, 56(5): 614-622.

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参考文献 36

[1]	He M, Zhou P, Chen H, et al. (-) -α-Bisabolol inhibits D-Gal-induced HSF cellular senescence in vitro and prevents skin aging in vivo by reducing SASP[J]. Iranian Journal of Basic Medical Sciences, 2024, 27 (9).
[2]	Umbayev B, Askarova S, Almabayeva A, et al. Galactose-induced skin aging: The role of oxidative stress[J]. Oxidative Medicine and Cellular Longevity, 2020: 7145656.
[3]	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 pmid: 31538262
[4]	Kumar H, Bhardwaj K, Valko M, et al. Antioxidative potential of Lactobacillus sp. in ameliorating D-galactose-induced aging[J]. Applied Microbiology and Biotechnology, 2022, 106 (13-16) : 4831-4843. doi: 10.1007/s00253-022-12041-7
[5]	Boismal F, Serror K, Dobos G, et al. Skin aging: Pathophysiology and innovative therapies[J]. Medecine Sciences, 2020, 36 (12) : 1163-1172.
[6]	Wang H, Sun Y, Qu T, et al. Nobiletin prevents D-galactose-induced C2C12 cell aging by improving mitochondrial function[J]. International Journal of Molecular Sciences, 2022, 23 (19) : 11963.
[7]	Zou X, Zhao M, Fei J, et al. Extraction, purification, structural identification, and anti-senescent activity of novel pearl peptides on human dermal fibroblasts[J]. Food Research International (Ottawa, Ont.), 2024, 198: 115357.
[8]	Elzi D J, Song M, Shiio Y. Role of galactose in cellular senescence[J]. Experimental Gerontology, 2016, 73: 1-4. doi: 10.1016/j.exger.2015.11.003 pmid: 26547052
[9]	王姝畅, 吴亚妮. 山茶籽油对D-半乳糖亚急性小鼠皮肤的影响[J]. 日用化学工业, 2019, 49 (5) : 320-327.
[10]	曹义苗, 高聪, 邓全知, 等. 高、低海拔红山茶花多酚类成分及抗皮肤老化功效研究[J]. 日用化学品科学, 2025, 48 (3) : 71-76.
[11]	曹义苗, 高宏旗, 高聪, 等. 高低海拔的山茶花活性成分及护肤功效研究[J]. 日用化学品科学, 2024, 47 (1) : 29-32.
[12]	曹义苗, 曲子越, 高聪, 等. 浙江红山茶不同组织活性成分及其抗氧化性研究[J]. 中国野生植物资源, 2024, 43 (8) : 41-45.
[13]	陈丹丹, 李广涛, 高宏旗, 等. 山茶花提取物功效研究[J]. 日用化学品科学, 2023, 46 (12) : 39-42, 48.
[14]	高宏旗, 李广涛, 陈斌, 等. 浙江红山茶种质资源、化学成分及生物活性研究进展[J]. 中国野生植物资源, 2024, 43 (S1) : 101-106.
[15]	胡昕, 李广涛, 高宏旗, 等. 红山茶花、叶提取液在化妆品中的抗老功效研究[J]. 中国野生植物资源, 2024, 43 (2) : 26-32.
[16]	李广涛, 高宏旗, 吕清爽, 等. 红山茶组合物诱导的人皮肤成纤维细胞自噬[J]. 香料香精化妆品, 2023 (6) : 110-115.
[17]	曲子越, 高宏旗, 曹义苗, 等. 油用山茶属植物的次生代谢成分及药理作用研究进展[J]. 香料香精化妆品, 2024 (2) : 41-50.
[18]	孙泽东, 李广涛, 王伊朋, 等. 山茶籽多肽制备及其抗老修护功效[J]. 精细化工, 2025, 42 (10) : 2232-2242.
[19]	李广涛, 孙泽东, 朱广宇, 等. 山茶籽油酶解前后理化性质和功效比较研究[J]. 中国野生植物资源, 2025, 44 (4) : 34-39, 47.
[20]	Ho S C, Liu J H, Wu R Y. Establishment of the mimetic aging effect in mice caused by D-galactose[J]. Biogerontology, 2003, 4 (1) : 15-18. doi: 10.1023/A:1022417102206
[21]	González-Casacuberta I, Juárez-Flores D L, Ezquerra M, et al. Mitochondrial and autophagic alterations in skin fibroblasts from Parkinson disease patients with Parkin mutations[J]. Aging, 2019, 11 (11) : 3750-3767. doi: 10.18632/aging.v11i11
[22]	Waggoner D D, Buist N R, Donnell G N. Long-term prognosis in galactosaemia: Results of a survey of 350 cases[J]. Journal of Inherited Metabolic Disease, 1990, 13 (6) : 802-818. pmid: 1706789
[23]	Asaduzzaman A K M, Getachew A T, Cho Y J, et al. Characterization of pepsin-solubilised collagen recovered from mackerel (Scomber japonicus) bone and skin using subcritical water hydrolysis[J]. International Journal of Biological Macromolecules, 2020, 148: 1290-1297. doi: S0141-8130(19)36244-0 pmid: 31739017
[24]	Chen H, Long Y, Guo L. Antiaging effect of Inula britannica on aging mouse model induced by D-galactose[J]. Evidence-Based Complementary and Alternative Medicine: eCAM, 2016: 6049083.
[25]	Pålsson-McDermott E M, O’Neill L A J. Targeting immunometabolism as an anti-inflammatory strategy[J]. Cell Research, 2020, 30 (4) : 300-314. doi: 10.1038/s41422-020-0291-z pmid: 32132672
[26]	Qin D, Ren R, Jia C, et al. Rapamycin protects skin fibroblasts from ultraviolet B-induced photoaging by suppressing the production of reactive oxygen species[J]. Cellular Physiology and Biochemistry, 2018, 46 (5) : 1849-1860. doi: 10.1159/000489369 pmid: 29705807
[27]	Dorf N, Maciejczyk M. Skin senescence-from basic research to clinical practice[J]. Frontiers in Medicine, 2024, 11: 1484345. doi: 10.3389/fmed.2024.1484345
[28]	Pan S, Gong S, Zhang J, et al. Anti-aging effects of fetal dermal mesenchymal stem cells in a D-galactose-induced aging model of adult dermal fibroblasts[J]. In Vitro Cellular & Developmental Biology - Animal, 2021, 57 (8) : 795-807. doi: 10.1007/s11626-021-00624-z
[29]	Mi W, Wang C, Luo G, et al. Targeting ERK induced cell death and p53/ROS-dependent protective autophagy in colorectal cancer[J]. Cell Death Discovery, 2021, 7 (1) : 375.
[30]	Chang Y Y, Wang M, Yeh J H, et al. The protective effects of beta-mangostin against sodium Iodate-induced retinal ROS-mediated apoptosis through MEK/ERK and p53 signaling pathways[J]. Food & Function, 2023, 14 (24) : 10896-10909.
[31]	Agostini F, Bisaglia M, Plotegher N. Linking ROS levels to autophagy: The key role of AMPK[J]. Antioxidants, 2023, 12 (7) : 1406.
[32]	Mihaylova M M, Shaw R J. The AMPK signalling pathway coordinates cell growth, autophagy and metabolism[J]. Nature Cell Biology, 2011, 13 (9) : 1016-1023. doi: 10.1038/ncb2329 pmid: 21892142
[33]	Xue C, Yan Z, Cheng W, et al. Curcumin ameliorates aging-induced blood-testis barrier disruption by regulating AMPK/mTOR mediated autophagy[J]. PLOS One, 2025, 20 (4) : e0321752.
[34]	Guo T, Liu Y, Ren X, et al. Promoting role of long non-coding RNA small nucleolar RNA host gene 15 (SNHG15) in neuronal injury following ischemic stroke via the microRNA-18a/CXC chemokine ligand 13 (CXCL13) /ERK/MEK axis[J]. Medical Science Monitor: International Medical Journal of Experimental and Clinical Research, 2020, 26.
[35]	Han Y, Wu N, Xia F, et al. Long noncoding RNA GAS5 regulates myocardial ischemiareperfusion injury through the PI3K/AKT apoptosis pathway by sponging miR5325p[J]. International Journal of Molecular Medicine, 2020, 45 (3) : 858-872.
[36]	Makrantonaki E, Bekou V, Zouboulis C C. Genetics and skin aging[J]. Dermato-Endocrinology, 2012, 4 (3) : 280-284. doi: 10.4161/derm.22372 pmid: 23467395

Gene	Forward primer	Reverse primer
CDKN2A（P16）	GATCCAGGTGGGTAGAAGGTC	CCCCTGCAAACTTCGTCCT
TP53（P53）	ACCTATGGAAACTACTTCCTGAAA	CTGGCATTCTGGGAGCTTCA
CDKN1A（P21）	CATGTGGACCTGTCACTGTC	TGGAGTGGTAGAAATCTGTCATGC
IL1-β	CAGAAGTACCTGAGCTCGCC	AGATTCGTAGCTGGATGCCG
IL-6	AGAGGCACTGGCAGAAAACA	CAGCTCTGGCTTGTTCCTCA
RELA（P65）	AACCTGGGAATCCAGTGTGTG	CCCACGCTGCTCTTCTTGGAA
GAPDH	ATTCCATGGCACCGTCAAGG	TCGCCCCACTTGATTTTGGA