人真皮成纤维细胞受长波紫外线辐照引起基因表达动态变化

doi:10.3969/j.issn.1001-1803.2021.09.010

摘要/Abstract

摘要：

通过不同累积剂量（1~7天）长波紫外线UVA辐照人原代真皮成纤维细胞,研究其细胞外基质、细胞周期及衰老相关基因动态变化情况。结果显示,胶原蛋白基因COL1A2、COL4A1在辐照1天时表达上调,辐照3天后下降;而弹性蛋白基因ELN在辐照1天时表达不变,辐照3天后下降;基质金属蛋白酶基因MMP1、MMP9在辐照5天时表达显著上调。UVA辐照3天和5天后,细胞阻滞于S期和G2/M期。细胞周期相关基因CDKN2A基因表达下调,TP53基因表达先上调后下调;CDKN1A基因表达上调,且在UVA辐照3天时上调幅度最大。衰老相关基因SIRT1、HDAC1与对照组相比,表现出先上调后下降,再上升的动态变化情况。结果表明UVA照射引发的细胞衰老变化是一个程序性调控的过程,成纤维细胞不同类型基因在UVA辐照下表现出差异的动态变化趋势,UVA辐照诱导细胞衰老模型应用于化妆品抗衰老功效研究时,应根据具体需求选择合适的生物学标志物。

关键词: 真皮成纤维细胞, 光衰老, 长波紫外线, 细胞外基质, 细胞周期, 化妆品

Abstract:

The dynamic changes of extracellular matrix, cell cycle and senescence-related genes at mRNA level in human primary dermal fibroblasts were studied by irradiated with different cumulative doses of ultraviolet UVA （1-7 days）. The results show that the expression of the collagen genes COL1A2 and COL4A1 is up-regulated at day 1 and decreases at day 3, while the mRNA level of the elastin gene ELN remains unchanged at day 1 and decreases at day 3. The expression of the matrix metalloproteinase genes MMP1 and MMP9 increases significantly at day 5 after UVA irradiation. Further studies show that the cell cycle is arrested in S phase and G2/M phase after 3 and 5 days after UVA irradiation. The expression of cell cycle related gene CDKN2A is down-regulated, and that of TP53 gene is up-regulated at first and then down-regulated. CDKN1A gene is up-regulated, and reaches the highest level at day 3 after UVA irradiation. Compared with the control group, the expression of aging-related genes SIRT1 and HDAC1 is up-regulated at first, then decreases, and increases again. The results show that the changes of cell senescence induced by UVA irradiation is a programmed regulation process, and different groups of genes in fibroblasts show diverse trends of dynamic changes under UVA irradiation. The suitable biomarkers should be selected according to the specific aims in the application of cell senescence model for researching anti-aging effect of cosmetics.

Key words: dermal fibroblasts, photoaging, ultraviolet A, extracellular matrix, cell cycle, cosmetics

中图分类号:

TQ658

殷庆飞,陈田. 人真皮成纤维细胞受长波紫外线辐照引起基因表达动态变化[J]. 日用化学工业, 2021, 51(9): 874-880.

Yin Qingfei,Chen Tian. Dynamic changes of gene expression induced by long-wave ultraviolet radiation in human dermal fibroblasts[J]. China Surfactant Detergent & Cosmetics, 2021, 51(9): 874-880.

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基因	方向	引物序列
COL1A2	F R	GTGGCAGTGATGGAAGTGTG AGGACCAGCGTTACCAACAG
COL4A1	F R	AGGTGTCATTGGGTTTCCTG GGTCCTCTTGTCCCTTTTGTT
ELN	F R	TCCAGGTGTAGGTGGAGCTT GTGTAGGGCAGTCCATAGCC
MMP1	F R	AAGGTGGACCAACAATTTCAGA TGAAGGTGTAGCTAGGGTACATCAA
MMP9	F R	CAGTCCACCCTTGTGCTCTT ATTTCGACTCTCCACGCATC
SIRT1	F R	CCCAGAACATAGACACGCTGGA TCAGCTGGGCACCTAGGACA
HDAC1	F R	ACCAAGTACCACAGCGATGACTACA CGAATACTGGACAGTCCTCACCAA
GDF11	F R	GTCATTAGCATGGCCCAGGA AGCTGGGCCTTCAGTACCTTTG
CDKN2A	F R	CCCCACTACCGTAAATGTCCA TGCTCACTCCAGAAAACTCCAA
TP53	F R	TCGAGATGTTCCGAGAGCTGAAT GTCTGAGTCAGGCCCTTCTGTCTT
CDKN1A	F R	CATGTGGACCTGTCACTGTCTTGTA GAAGATCAGCCGGCGTTTG
ACTB	F R	TGGCACCCAGCACAATGAA CTAAGTCATAGTCCGCCTAGAAGCA