藏绵羊瘤胃上皮ceRNA介导GPCR-cAMP-PKA信号通路调控SCFAs  

Regulation of SCFAs via GPCR-cAMP-PKA Signaling Pathway Mediated by ceRNA in Tibetan Sheep Rumen Epithelium

作　　者：沙玉柱 刘秀 赵生国 邵鹏阳 王翻兄 杨文鑫 陈小伟 谢转回 SHA Yuzhu;LIU Xiu;ZHAO Shengguo;SHAO Pengyang;WANG Fanxiong;YANG Wenxin;CHEN Xiaowei and XIE Zhuanhui(College of Animal Science and Technology,Gansu Agricultural University/Gansu Key Laboratory of Herbivore Biotechnology,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院/甘肃省草食动物生物技术重点实验室,兰州730070

出　　处：《西北农业学报》2025年第4期581-591,共11页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(32260820);甘肃农业大学科技创新基金(GAU-QDFC-2022-06);甘肃海智计划示范项目(GSHZSF2023-01);甘肃农业大学学科团队项目(GAU-XKTD-2022-21)。

摘　　要：旨在探究藏绵羊瘤胃上皮对短链脂肪酸(SCFAs)的吸收转运分子机制。选择1周岁(±1月龄)健康的放牧藏绵羊母羊12只,分别对其暖季(8月份)和冷季(12月份)的瘤胃上皮组织进行全转录组(RNAseq)测序,筛选差异ceRNA调控网络,检测cAMP-PKA信号通路基因的表达量,并分析其与瘤胃微生物菌群及其代谢物SCFAs的相关性。结果表明:冷、暖季藏绵羊瘤胃上皮mRNA和非编码RNA(miRNA/circRNA)表达量存在差异,筛选到差异ceRNA调控网络AKAP9-circRNA(NC_040269.1:68309959|68364662)-miR-370-3p;靶基因AKAP9的GO功能富集分析(Gene Ontology Enrichment Analysis)发现主要富集在钾离子跨膜转运蛋白活性的正调控、信号转导和细胞对cAMP的反应等通路中。分析cAMP-PKA信号通路相关基因的表达特征,发现PKA表达量在冷、暖季差异不显著(P>0.05),与PKA锚定的AKAP9在冷季表达量显著升高(P<0.05),而G蛋白偶联受体GPR41在冷季表达量显著降低(P<0.05)。相关性分析发现,乙酸、丙酸与AKAP9和GPR41呈极显著性正相关(P<0.01),异丁酸与CREB1呈显著性负相关(P<0.05),瘤胃微生物菌群与SCFAs存在显著相关性(P<0.05)。在冷季营养胁迫下,藏绵羊瘤胃上皮非编码RNA通过ceRNA网络参与调控cAMP-PKA信号通路,进而影响瘤胃上皮SCFAs的吸收转运。Twelve healthy Tibetan ewes,aged 1 year(±1 month),were selected to investigate the absorption and transport mechanism of short-chain fatty acids(SCFAs)in the rumen epithelium of Tibetan sheep.Rumen epithelial tissue samples were collected during both cold and warm seasons for whole transcriptome sequencing(RNA-seq).Differential ceRNA regulatory networks were identified.The expression levels of genes related to the cAMP-PKA signaling pathway were analyzed.The correlation between cAMP-PKA signaling pathway genes and rumen microbes as well as SCFAs was examined.Tibetan sheep rumen epithelial mRNA and non-coding RNA(miRNA/circRNA)expression quantity differences in cold and warm seasons,and a ceRNA regulation network involving AKAP9-circRNA(NC_040269.1:68309959|68364662)-miR-370-3p were selected for analysis.GO functional enrichment analysis was performed on the target gene AKAP9,it revealed enrichment in positive regulation of potassium ion transmembrane transporter activity,signal transduction,and cellular response to the cAMP pathway.Further analysis of gene expression related to the cAMP-PKA signaling pathway revealed that PKA did not show significant differences between the cold and warm seasons(P>0.05).However,the expression of AKAP9,which binds to PKA,significantly increased during the cold season(P<0.05),while the expression of the G-protein coupled receptor GPR41 showed a significant decrease during the cold season(P<0.05).Correlation analysis revealed a positive correlation between acetic acid and propionic acid with both AKAP9 and GPR41(P<0.01).Additionally,a significant negative correlation was observed between isobutyrate and CREB1(P<0.05).Furthermore,a significant correlation was identified between the ruminal microbiota and SCFAs.The non-coding RNA in the rumen epithelium of Tibetan sheep participates in the regulation of the cAMP-PKA signaling pathway through the ceRNA network.This process influences the absorption and transport of SCFAs in the rumen.

关 键 词：冷季 藏绵羊 SCFAs 瘤胃上皮 ceRNA cAMP 

分 类 号：S826[农业科学—畜牧学]