不同氧浓度下猪睾丸间质细胞转录组分析

doi:10.16431/j.cnki.1671-7236.2023.06.002

摘要/Abstract

摘要： 【目的】研究不同氧浓度下猪睾丸间质细胞表达谱差异，筛选缺氧导致猪睾丸间质细胞损伤的关键基因。【方法】将猪睾丸间质细胞分为缺氧组（1%氧气，H24组）、正常组（15.75%氧气，N24组），每组3个重复，在相应条件下分别处理24 h，用CCK8法检测细胞存活率；利用转录组测序技术筛选出缺氧组和正常组睾丸间质细胞中的差异表达基因，利用DESeq 2.0软件进行差异表达基因分析，并对差异表达基因进行GO功能和KEGG通路富集分析，进一步筛选与氧含量相关的调控基因。【结果】缺氧刺激24 h后，与正常组相比，缺氧组猪睾丸间质细胞存活率极显著降低（P<0.01）；转录组测序共获得1 654个差异表达基因，其中1 242个基因上调，412个基因下调。GO功能富集分析发现，共获得富集条目8条，其中生物过程1条，分子功能7条；KEGG通路富集分析发现，共获得10个显著富集的信号通路，包括HIF-1信号通路、JAK-STAT信号通路和糖尿病并发症中的AGE-RAGE信号通路等。通过GO功能和KEGG通路富集分析共获得6个与缺氧应激相关的基因，分别为信号传导及转录激活蛋白5（signal transducer and activator of transcription 5，STAT5）、白细胞介素-8（interleukin-8，IL-8）、CC基元趋化因子配体2（C-C motif chemokine ligand 2，CCL2）、Ⅰ型干扰素受体1（interferon alpha and beta receptor subunit 1，IFNAR1）、细胞因子信号传导抑制因子1（suppressor of cytokine signaling 1，SOCS1）和红细胞生成素（erythropoietin，EPO）。【结论】本研究通过对不同氧浓度下猪睾丸间质细胞进行转录组分析，获得了SOCS1、IFNAR1、EPO、STAT5、IL-8和CCL2 6个与猪睾丸间质细胞缺氧损伤和应激修复相关的基因，为解决高原地区公猪生殖能力低下的研究提供参考。

关键词: 缺氧; 猪睾丸间质细胞; 生殖损伤; 转录组测序

Abstract: 【Objective】 The purpose of this study was to explore the difference in the expression profile of porcine Leydig cells under different oxygen concentrations and screen the key genes of hypoxia leading to the damage of porcine Leydig cells.【Method】 Porcine Leydig cells were divided into hypoxia group (1% oxygen, H24 group) and normal group (15.75% oxygen, N24 group) with 3 replicates in each group.The cells were treated for 24 h under corresponding conditions, and the cell survival rate was detected by CCK8 method.Transcriptome sequencing technology was used to screen out the differentially expressed genes in Leydig cells of hypoxia and normal oxygen groups.DESeq 2.0 software was used to analyze the differentially expressed genes, and GO function and KEGG pathway enrichment analysis were performed on the differentially expressed genes, so as to further screen the regulatory genes related to oxygen content.【Result】 After 24 h hypoxia stimulation, the survival rate of porcine Leydig cells in hypoxia group was extremely significantly decreased compared with normal group (P<0.01).A total of 1 654 differentially expressed genes were obtained by transcriptome sequencing, among which 1 242 genes were up-regulated and 412 genes were down-regulated.GO function enrichment analysis showed that 8 enrichment items were obtained, including 1 biological process and 7 molecular functions.KEGG pathway enrichment analysis showed that 10 signaling pathways were significantly enriched, including HIF-1 signaling pathway, JAK-STAT signaling pathway and AGE-RAGE signaling pathway in diabetic complications, etc.Six genes related to hypoxia stress were obtained by enrichment analysis of GO function and KEGG pathway, including signal transducer and activator of transcription 5(STAT5), interleukin-8 (IL-8), C-C motif chemokine ligand 2 (CCL2), interferon alpha and beta receptor subunit 1 (IFNAR1), suppressor of cytokine signaling 1 (SOCS1) and erythropoietin (EPO).【Conclusion】 In this study, six genes related to hypoxia injury and stress repair in porcine Leydig cells (SOCS1, IFNAR1, EPO, STAT5, IL-8 and CCL2 genes) were obtained through transcriptomic analysis of porcine Leydig cells under different oxygen concentrations, providing reference for the study on the low reproductive ability of boar in plateau area.

Key words: hypoxia; porcine Leydig cells; reproductive injury; transcriptome sequencing

中图分类号:

S828

陈楷文, 赵云娇, 王磊, 张剑搏, 马玉红, 吴国芳. 不同氧浓度下猪睾丸间质细胞转录组分析[J]. 中国畜牧兽医, 2023, 50(6): 2185-2195.

CHEN Kaiwen, ZHAO Yunjiao, WANG Lei, ZHANG Jianbo, MA Yuhong, WU Guofang. Transcriptome Analysis of Porcine Leydig Cells Under Different Oxygen Concentrations[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2185-2195.

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