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  

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

作　　者：廖峰 樊雨梅 帖航 赵海晴 苏宁[2] LIAO Feng;FAN Yumei;TIE Hang;ZHAO Haiqing;SU Ning(National Engineering Research Center for Gelatin-based Traditional Chinese Medicine,Dong-E-E-Jiao Co.,Ltd.,Liaocheng 252201,China;Chinese Academy of Inspection and Quarantine,Beijing 100176,China)

机构地区：[1]国家胶类中药工程技术研究中心,东阿阿胶股份有限公司,山东聊城252201 [2]中国检验检疫科学研究院,北京100176

出　　处：《食品工业科技》2022年第6期358-364,共7页Science and Technology of Food Industry

基　　金：山东省泰山产业领军人才传统产业创新类项目(TSCY20170233)。

摘　　要：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,驴骨胶原蛋白对NHDFs细胞无毒性,且在0.16~5.0 g/mL内对NHDFs细胞有一定的促生长作用。0.75~1.50 g/mL的驴骨胶原蛋白能够促进NHDFs细胞增殖,且驴骨胶原蛋白对NHDFs细胞增殖的影响呈现剂量依赖关系。1.50 g/mL的驴骨胶原蛋白可显著促进NHDFs细胞迁移和Ⅰ胶原蛋白的分泌。驴骨胶原蛋白作用于成纤维细胞后,共筛选出384个差异表达基因,其中172个基因上调,212个基因下调。GO功能富集分析结果表明,差异表达基因与中性粒细胞趋化性、中性粒细胞迁移、粒细胞趋化性等有关;KEGG通路富集分析表明,差异基因参与细胞因子-细胞因子受体相互作用、化学致癌、IL-17和TNF等重要生物学通路。推测,驴骨胶原蛋白可能通过IL-17和TNF-α介导CXCL趋化因子发挥促进皮肤伤口愈合的作用。本文明确了驴骨胶原蛋白促进伤口愈合的效果,填补驴骨胶原蛋白促进伤口愈合的研究空白,为驴骨胶原蛋白高值转化提供了思路。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 downregulated. 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.

关 键 词：驴骨胶原蛋白 伤口愈合 NHDFs细胞 Ⅰ型胶原蛋白 RNA-seq技术 

分 类 号：TS251.9[轻工技术与工程—农产品加工及贮藏工程]