不同年龄阶段天祝白牦牛皮肤转录组分析及免疫基因筛选

doi:10.16431/j.cnki.1671-7236.2025.08.002

摘要/Abstract

摘要： 【目的】通过对不同年龄阶段天祝白牦牛肩胛部皮肤组织进行转录组测序，筛选不同年龄阶段的差异表达基因(DEGs)，分析DEGs的功能和互作关系，为解析牦牛皮肤组织发育过程适应高寒环境的分子机理提供理论依据。【方法】采集0.5、2.5、4.5岁3个年龄阶段体态健康、生长状况良好的天祝白牦牛采集肩胛部皮肤进行转录组测序，以Q＜0.05、|log₂FoldChange|＞1为阈值筛选DEGs，并对其进行GO功能和KEGG通路富集分析。随机挑选7个DEGs进行实时荧光定量PCR验证转录组测序结果。【结果】在获得的605个DEGs中，2.5 vs 0.5、4.5 vs 0.5、4.5 vs 2.5比较组分别筛选出364、153、88个DEGs。GO功能富集分析显示，在2.5 vs 0.5比较组中DEGs与免疫反应相关的有积极调节免疫反应、角质化、中性粒细胞趋化以及趋化因子活性；在4.5 vs 0.5比较组中DEGs与免疫反应相关的有趋化因子正调控分泌、趋化因子活性以及对细菌的反应；在4.5 vs 2.5比较组中DEGs与免疫反应相关的有免疫反应、肽酶抑制剂活性。KEGG通路注释结果显示，在2.5 vs 0.5、4.5 vs 0.5比较组中DEGs均显著富集在病毒蛋白与细胞因子及细胞因子受体的相互作用、IL-17信号通路、细胞因子-细胞因子受体相互作用等免疫相关通路；在4.5 vs 2.5比较组中DEGs显著富集在ABC转运蛋白、金黄色葡萄球菌感染、补体和凝血级联反应。通过富集结果筛选出与皮肤免疫调节相关的30个基因，利用STRING数据库及Cytoscape软件制作蛋白互作网络分析图，并筛选出了IL-6、IL-1A、CCL20 3个得分较高的蛋白作为核心蛋白。实时荧光定量PCR结果与转录组测序表达趋势一致，表明转录组测序结果可靠。【结论】本研究揭示了不同年龄阶段天祝白牦牛皮肤组织中DEGs数量，以及DEGs的功能和所在通路，其中IL-17信号通路等与天祝白牦牛皮肤免疫功能发育及调控有关，筛选出IL-6、IL-1A、CCL20等关键调控基因。研究结果为全面了解天祝白牦牛皮肤免疫功能调控机制及相关基因提供了参考。

关键词: 天祝白牦牛; 皮肤; 转录组; 免疫

Abstract: 【Objective】 The transcriptome sequencing was performed on the skin tissue of the scapular region in Tianzhu White yaks at different age stages,the differentially expressed genes (DEGs) at different age stages were screened,and the function and interaction relationship of DEGs were analyzed,so as to provide a theoretical basis for analyzing the molecular mechanism of skin tissue development process in yak adapting to the alpine environment. 【Method】 The skin tissue of the scapular region in Tianzhu White yaks in good health and growth condition at the ages of 0.5,2.5,and 4.5 years were collected for RNA-Seq.The DEGs were screened with the thresholds of Q＜0.05 and |log₂FoldChange|＞1.Then,GO function and KEGG pathway enrichment analysis were conducted for DEGs.Seven DEGs were randomly selected for Real-time quantitative PCR to verify RNA-Seq results. 【Result】 Among the 605 obtained DEGs,364,153,and 88 DEGs were screened in 2.5 vs 0.5,4.5 vs 0.5,and 4.5 vs 2.5 groups.GO functional enrichment analysis showed that DEGs in 2.5 vs 0.5 group were related to the immune response included positive regulation of immune response,keratinization,neutrophil chemotaxis and chemokine activity,DEGs in 4.5 vs 0.5 group were related to the immune response included positive regulation of chemokine secretion,chemokine activity and response to bacteria,and DEGs in 4.5 vs 2.5 group were related to the immune response included immune response and peptidase inhibitor activity.KEGG pathway annotation results indicated that DEGs in 2.5 vs 0.5 and 4.5 vs 0.5 groups were significantly enriched in immune-related pathways such as the interaction between viral proteins and cytokines and cytokine receptors,IL-17 signaling pathway,and cytokine-cytokine receptor interaction,and DEGs in 4.5 vs 2.5 group were significantly enriched in ABC transporters, Staphylococcus aureus infection,complement and coagulation cascade reaction,etc.Through the enrichment results,30 genes related to skin immune regulation were screened out,and the protein-protein interaction network analysis map was made by STRING database and Cytoscape software,and IL-6,IL-1A,and CCL20 with higher scores were screened as the core proteins.The results of Real-time quantitative PCR were consistent with the expression trend of RNA-Seq,indicating that RNA-Seq results were reliable. 【Conclusion】 The number of DEGs in skin tissues of Tianzhu White yaks at different age stages were revealed in this study,as well as the function and pathway of DEGs.Through analysis,it was believed that pathways such as the IL-17 signaling pathway were associated with the development and regulation of the skin immune function in Tianzhu White yaks,and key regulatory genes such as IL-6,IL-1A and CCL20 were screened.The results provided a reference for comprehensively understanding the regulation mechanism of the skin immune function and related genes in Tianzhu White yaks.

Key words: Tianzhu White yak; skin; transcriptome; immunity

中图分类号:

S823.8⁺5

邓赟奇, 石斌刚, 赵志东, 胡江, 张哓兰. 不同年龄阶段天祝白牦牛皮肤转录组分析及免疫基因筛选[J]. 中国畜牧兽医, 2025, 52(8): 3515-3526.

DENG Yunqi, SHI Bingang, ZHAO Zhidong, HU Jiang, ZHANG Xiaolan. Transcriptome Analysis of Tianzhu White Yak Skin at Different Age Stages and Identification of Immune-regulating Genes[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3515-3526.

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