高通量测序解析驴骨胶原蛋白促进伤口愈合的潜在分子机制

探讨驴骨胶原蛋白对成纤维细胞(NHDFs细胞)增殖、迁移和I型胶原蛋白分泌的影响以及可能的作用通路.通过八肽胆囊收缩素(CCK-8)法、5-溴脱氧尿嘧啶核苷(BrdU)法和划痕实验,研究驴骨胶蛋白对成纤维细胞增殖和迁移的影响,并利用转录组测序技术(RNA-seq)分析驴骨胶原蛋白作用于成纤维细胞的差异表达基因.结果表明...

Analysis of the Potential Molecular Mechanism of Donkey Bone Collagen Promoting Skin Wound Healing by High-throughput Sequencing

To explore the effects of donkey bone collagen on the proliferation, migration and secretion of type I collagen of fibroblasts (NHDFs) and its possible signal pathways, the effects of donkey bone collagen on the proliferation and migration of fibroblasts were studied by CCK-8, BrdU and scratch test, and the differentially expressed genes of donkey bone collagen acting on fibroblasts were analyzed by RNA-seq. The results showed that donkey bone collagen had no toxicity to NHDFs cells, and could promote the growth of NHDFs cells within 0.16~5.0 g/mL. 0.75~1.50 g/mL donkey bone collagen could promote the proliferation of NHDFs cells, and the effect of donkey bone collagen on the proliferation of NHDFs cells showed a dose-dependent relationship. 1.50 g/mL donkey bone collagen could significantly promote the migration of NHDFs cells and the secretion of type I collagen. After donkey bone collagen acted on fibroblasts, 384 differentially expressed genes were screened out, of which 172 genes were up-regulated and 212 genes were down-regulated. The results of GO functional enrichment analysis showed that differentially expressed genes were related to neutrophil chemotaxis, neutrophil migration and granulocyte chemotaxis. Enrichment analysis of KEGG pathway showed that differential genes were involved in important biological pathways such as cytokine-cytokine receptor interaction, chemical carcinogenesis, IL-17 and TNF signaling pathway. It was speculated that donkey bone collagen may promote skin wound healing through IL-17 and TNF-α-mediated CXCL chemokines. It clarified the effect of donkey bone collagen in promoting wound healing, filled the gap in the research of donkey bone collagen in promoting wound healing, and would provide ideas for the high-value transformation of donkey bone collagen.