基于转录组学研究双酚A对猪睾丸支持细胞炎症和氨基酸代谢通路的影响  

The Effects of Bisphenol A on Inflammation and Amino Acid Metabolism Pathways in Porcine Testis Sertoli Cells Based on Transcriptome Analysis

作　　者：胡婷 张永红[1] 侯晓林[1] 姚华[1] 崔德凤[1] 潘早早 张凌宇 张家希 吴琼 HU Ting;ZHANG Yonghong;HOU Xiaolin;YAO Hua;CUI Defeng;PAN Zaozao;ZHANG Lingyu;ZHANG Jiaxi;WU Qiong(Animal Science and Technology College,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院动物科学技术学院,北京102206

出　　处：《畜牧兽医学报》2023年第7期2858-2871,共14页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32202881)。

摘　　要：旨在研究猪睾丸支持细胞(ST)暴露于环境雌激素双酚A(BPA)不同时间后的基因表达谱变化。本研究将猪ST细胞设为C、B两组,每组设置3个重复,C组为空白对照组,B组细胞暴露于50μmol·L^(-1)浓度的BPA。BPA暴露6、24 h后,收集C6、B6、C24、B24组细胞总RNA样品,构建测序文库,通过双端150 bp测序方法进行高通量转录组测序。对组装数据进行功能注释、差异基因分析以及GO和KEGG富集分析。通过Real-time PCR方法对关键差异基因表达进行验证。BPA暴露24 h后差异基因数量(6928个)较BPA暴露6 h差异基因数量(3940个)明显增加。编码分泌型磷蛋白1的SPP1基因在BPA暴露6 h后表达增加,但在BPA暴露24 h后表达降低。管腔结合蛋白编码基因BIP、泛素B编码基因UBB、泛素C编码基因UBC、鸟氨酸脱羧酶1编码基因ODC1表达量在BPA暴露6和24 h后均显著上调。富集分析结果表明,BPA暴露6 h后,TNF信号通路富集差异倍数最高,p53、IL-17以及MAPK信号通路等也均显著富集,暗示ST细胞表现出明显的促炎反应。在BPA暴露24 h后,氨基酸及糖代谢通路富集差异倍数最高,包括精氨酸和脯氨酸、组氨酸以及糖酵解代谢通路。Real-time PCR验证结果显示,BPA暴露6 h后炎性反应通路相关的PTGS2等以及BPA暴露24 h后氨基酸代谢通路相关ARG1等差异基因表达显著上调。综上可知,BPA对猪睾丸支持细胞的损伤机制呈现明显的时间效应,ST细胞在BPA暴露初期表现出以TNF等信号通路激活为特征的炎性反应,在BPA暴露后期氨基酸及糖代谢显著激活。This experiment was conducted to compare the gene expression profile changes of porcine testicular sertoli cells(ST)exposed to environmental estrogen bisphenol A(BPA)for different times.The porcine ST cells were divided into group C and group B with 3 replicates per group.Group C was a blank control group,and group B cells were exposed to BPA at a concentration of 50μmol·L^(-1).At 6 and 24 h after BPA exposure,total RNA samples from C 6,B 6,C 24 and B 24 group cells were collected,and a sequencing library was constructed.High-throughput transcriptome sequencing was performed using a paired-end 150 bp sequencing method.The assembly data were used for functional annotation,differentially expressed gene(DEG)analysis,and GO and KEGG enrichment analysis.The expression of some key DEGs were validated through real-time PCR method.The number of DEGs(6928)significantly increased at 24 h after BPA exposure,compared with 3940 DEGs at 6 h.The expression of SPP 1 gene encoding secretory phosphoprotein 1 increased at 6 h after BPA exposure,but decreased at 24 h after BPA exposure.The expression of lumen binding protein-encoding gene BIP,ubiquitin B-encoding gene UBB,ubiquitin C-encoding gene UBC,and ornithine decarboxylase 1-encoding gene ODC 1 were significantly upregulated both at 6 and 24 h after BPA exposure.Enrichment analysis showed that the TNF signal pathway was the most significantly enriched at 6 h,and p53,IL-17 and MAPK signal pathways were also significantly enriched,indicating that ST cells exhibited proinflammatory response.At 24 h after BPA exposure,DEGs in pig ST cells were enriched in amino acids and sugar metabolism pathways,including arginine and proline,histidine and glycolysis metabolic pathways.Real-time PCR results showed that PTGS 2 and other DEGs related to the inflammatory response pathway at 6 h after BPA exposure and ARG 1 and other DEGs related to the amino acid metabolic pathways at 24 h after BPA exposure were significantly up-regulated.In conclusion,the damage mechanism of BPA on porcine serto

关 键 词：双酚A 猪睾丸支持细胞 转录组测序 富集分析 

分 类 号：S828.3[农业科学—畜牧学]