Fang Xiaokai, Zhang Shan, Wu Mingyang, Luo Yang, Chen Xingyu, Zhou Yuan, Zhang Yu, Liu Xiaochun, Yao Xu
Department of Allergy and Rheumatology, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Hospital for Skin Diseases, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing, Jiangsu 210042, China.
Chin Med J (Engl). 2025 Sep 5;138(17):2180-2191. doi: 10.1097/CM9.0000000000003312. Epub 2024 Sep 27.
Senescent human skin primary fibroblast (FB) models have been established for studying aging-related, proliferative, and inflammatory skin diseases. The aim of this study was to compare the transcriptome characteristics of human primary dermal FBs from children and the elderly with four senescence models.
Human skin primary FBs were obtained from healthy children (FB-C) and elderly donors (FB-E). Senescence models were generated by ultraviolet B irradiation (FB-UVB), D-galactose stimulation (FB-D-gal), atazanavir treatment (FB-ATV), and replication exhaustion induction (FB-P30). Flow cytometry, immunofluorescence staining, real-time quantitative polymerase chain reaction, co-culturing with immune cells, and bulk RNA sequencing were used for systematic comparisons of the models.
In comparison with FB-C, FB-E showed elevated expression of senescence-related genes related to the skin barrier and extracellular matrix, proinflammatory factors, chemokines, oxidative stress, and complement factors. In comparison with FB-E, FB-UVB and FB-ATV showed higher levels of senescence and expression of the genes related to the senescence-associated secretory phenotype (SASP), and their shaped immune microenvironment highly facilitated the activation of downstream immune cells, including T cells, macrophages, and natural killer cells. FB-P30 was most similar to FB-E in terms of general transcriptome features, such as FB migration and proliferation, and aging-related characteristics. FB-D-gal showed the lowest expression levels of senescence-related genes. In comparisons with the single-cell RNA sequencing results, FB-E showed almost complete simulation of the transcriptional spectrum of FBs in elderly patients with atopic dermatitis, followed by FB-P30 and FB-UVB. FB-E and FB-P30 showed higher similarity with the FBs in keloids.
Each senescent FB model exhibited different characteristics. In addition to showing upregulated expression of natural senescence features, FB-UVB and FB-ATV showed high expression levels of senescence-related genes, including those involved in the SASP, and FB-P30 showed the greatest similarity with FB-E. However, D-galactose-stimulated FBs did not clearly present aging characteristics.
已建立衰老的人皮肤原代成纤维细胞（FB）模型，用于研究与衰老相关的、增殖性和炎症性皮肤病。本研究的目的是比较来自儿童和老年人的人原代真皮成纤维细胞与四种衰老模型的转录组特征。
从健康儿童（FB-C）和老年供体（FB-E）获取人皮肤原代成纤维细胞。通过紫外线B照射（FB-UVB）、D-半乳糖刺激（FB-D-gal）、阿扎那韦处理（FB-ATV）和复制耗竭诱导（FB-P30）生成衰老模型。采用流式细胞术、免疫荧光染色、实时定量聚合酶链反应、与免疫细胞共培养以及大量RNA测序对模型进行系统比较。
与FB-C相比，FB-E中与皮肤屏障和细胞外基质、促炎因子、趋化因子、氧化应激及补体因子相关的衰老相关基因表达升高。与FB-E相比，FB-UVB和FB-ATV表现出更高水平的衰老以及与衰老相关分泌表型（SASP）相关基因的表达，且它们形成的免疫微环境极大地促进了包括T细胞、巨噬细胞和自然杀伤细胞在内的下游免疫细胞的激活。FB-P30在一般转录组特征（如FB迁移和增殖）以及与衰老相关的特征方面与FB-E最为相似。FB-D-gal显示衰老相关基因的表达水平最低。与单细胞RNA测序结果比较，FB-E几乎完全模拟了特应性皮炎老年患者成纤维细胞的转录谱，其次是FB-P30和FB-UVB。FB-E和FB-P30与瘢痕疙瘩中的成纤维细胞表现出更高的相似性。
每个衰老的FB模型都表现出不同的特征。除了显示自然衰老特征的表达上调外，FB-UVB和FB-ATV还显示出与衰老相关基因（包括参与SASP的基因）的高表达，且FB-P30与FB-E表现出最大的相似性。然而，D-半乳糖刺激的成纤维细胞并未明显呈现衰老特征。
