Regulatory role of ADAMTS8 in proliferation and apoptosis of hypertrophic scar fibroblasts

doi:10.12307/2026.811

Abstract

Abstract: BACKGROUND: Studies have confirmed that A disintegrin and metalloproteinase with thrombospondin motifs 8 (ADAMTS8) plays a regulatory role in fibrosis. Therefore, elucidating the mechanism of ADAMTS8 in hypertrophic scar formation holds considerable clinical significance.
OBJECTIVE: To investigate the regulatory role of ADAMTS8 in scar fibroblasts.
METHODS: (1) Immunohistochemical staining was used to detect the expression of type I collagen, type III collagen, α-smooth muscle actin, and ADAMTS8 in normal human skin tissue and hypertrophic scar tissue. The protein expression level of ADAMTS8 was detected by western blot assay in normal human skin tissue and hypertrophic scar tissue. Hypertrophic scar tissue was taken as positive sample and normal human skin tissue was taken as negative sample. Receiver operating characteristic curve was drawn to analyze the ability of ADAMTS8 to predict the difference between normal human skin and hypertrophic scar. (2) A protein-protein interaction network of ADAMTS8 was constructed using the STRING 12.0 platform, and Gene Ontology function enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted on the obtained targets. (3) Human hypertrophic scar tissue fibroblasts were isolated and cultured. The 3rd to 6th generation fibroblasts were selected and divided into three groups: the control group was cultured routinely; empty vector group (Ad-NC) transfected with empty vector adenovirus, and overexpressing ADAMTS8 group (Ad-ADAMTS8) was transfected with ADAMTS8 overexpressing adenovirus. The cell proliferation activity was detected by CCK8 assay and EDU staining. Cell apoptosis was detected by flow cytometry and TUNEL staining.
RESULTS AND CONCLUSION: (1) Immunohistochemical staining demonstrated significantly higher protein expression of type I collagen, type III collagen, and α-smooth muscle actin in hypertrophic scars compared with normal skin (P < 0.001). In contrast, ADAMTS8 expression was significantly lower in hypertrophic scars (P < 0.001). Western blot assay showed that ADAMTS8 protein expression was significantly lower in hypertrophic scar tissue than in normal skin (P < 0.001). Receiver operating characteristic curve analysis indicated that the area under the curve for ADAMTS8 in predicting the development of hypertrophic scars was 0.86, indicating its good ability to discriminate between hypertrophic scar and normal skin. (2) The top 41 genes were screened using STRING database. Kyoto Encyclopedia of Genes and Genomes enrichment showed that ADAMTS8 was mainly involved in the biological processes and key mechanisms such as extracellular matrix receptor interaction, phosphatidylinositol-3-kinase-protein kinase B signaling pathway and efferocytosis. Gene ontology enrichment showed that ADAMTS8 was involved in apoptosis-related pathway enrichment, including negative regulation of fibroblast growth factor receptor signaling pathway, binding of fibroblast growth factor, negative regulation of apoptosis and apoptosis process. (3) CCK-8 assay and EdU staining showed that of ADAMTS8 overexpression could inhibit the proliferation of hypertrophic scar fibroblasts. Flow cytometry and TUNEL staining showed that ADAMTS8 overexpression could promote the apoptosis of hypertrophic scar fibroblasts. (4) These results indicate that ADAMTS8 expression is significantly decreased in human hypertrophic scars, and ADAMTS8 overexpression can inhibit proliferation of hypertrophic scar fibroblasts and promote apoptosis of hypertrophic scar fibroblasts.

Key words: hypertrophic scar, ADAMTS8, fibrosis, fibroblast, proliferation, apoptosis, tissue construction

CLC Number:

Zhang Jingyi, Ma Fang, Liu Honglin, Wang Jianjun, Xia Tongtong, Yang Jiaqi, Wang Yajing, Shen Jiangyong, Jiang Yideng. Regulatory role of ADAMTS8 in proliferation and apoptosis of hypertrophic scar fibroblasts[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(28): 7260-7266.

Figures/Tables 5

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