ploy(I∶C)刺激马氏珠母贝血淋巴外泌体miRNA差异表达分析  

Identification of differentially expressed miRNAs in haemolymph exosomes after poly(I∶C)stimulation in Pinctada fucata martensii

作　　者：王萍 谭婉琦 任紫绮 李芷晴 左贤 屈佳 王庆恒[1,2,3,4] 郑哲 WANG Ping;TAN Wanqi;REN Ziqi;LI Zhiqing;ZUO Xian;QU Jia;WANG Qingheng;ZHENG Zhe(Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Science and Innovation Center for Pearl Culture,Zhanjiang 524088,China;Guangdong Pearl Cultivation and Processing Engineering and Technology Research Centre,Zhanjiang 524088,China;Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture,Zhanjiang 524088,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东省珍珠科技创新中心,广东湛江524088 [3]广东省珍珠养殖与加工工程技术研究中心,广东湛江524088 [4]广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088

出　　处：《中国水产科学》2024年第11期1375-1386,共12页Journal of Fishery Sciences of China

基　　金：湛江市科技计划项目(2022A01010);广东海洋大学科研项目经费(060302022305);广东海洋大学创新创业训练计划项目(CXXL2022011);广东省自然科学基金项目(2023A1515030048,2024A1515011617)

摘　　要：外泌体mi RNAs在调控脊椎动物的免疫应答过程中发挥重要作用。为探究外泌体mi RNA在马氏珠母贝免疫应答过程中的作用,通过向马氏珠母贝闭壳肌血窦中注射poly (I∶C)和PBS进行免疫刺激,分析血淋巴中差异表达的外泌体mi RNAs,预测其靶基因及调控的免疫相关信号通路。结果显示,马氏珠母贝血淋巴外泌体为球形或近似球形的囊泡,膜结构完整、轮廓清晰,粒径为75~150 nm。PBS组和poly (I∶C)组共鉴定到101条mi RNA,其中76条为已知mi RNA。与PBS组相比,poly (I∶C)组有7条mi RNA显著上调,5条mi RNA显著下调。以上差异mi RNA共获得570个靶基因。KEGG富集分析发现了与免疫相关的信号通路包括“acute myeloid leukemia”“JAK-STAT signaling pathway”和“chemokine signaling pathway”,与代谢相关的“glycerolipid metabolism”“purine metabolism”和“arginine and proline metabolism”以及与内分泌相关的“relaxin signaling pathway”“prolectin signaling pathway”和“insect hormone biosynthesis”(P<0.05)。其中,novel_mir2、novel_mir22、mi R-2b_5、mi R-279等8个mi RNAs在免疫相关的信号通路中发挥调控作用,并通过FOXO偶联调控代谢影响马氏珠母贝的免疫应答。以上研究表明马氏珠母贝血淋巴外泌体mi RNA通过调控免疫、代谢和内分泌等协调响应poly (I∶C)刺激,结果为深入研究贝类中外泌体mi RNA在免疫应答中的作用提供基础资料。Exosomal miRNAs are crucial in regulating the immune response of vertebrates.This study explored the function of exosomal miRNAs in the immune response of Pinctada fucata martensii by injecting poly(I∶C)and PBS into the blood sinuses of the adductor muscle of P.f.martensii to stimulate an immune response,followed by the analysis of differentially expressed exosomal miRNAs in the hemolymph.The study aimed to predict their target genes and the immune-related signaling pathways regulated by exosomal miRNAs,providing preliminary insights into their roles in the immune response of P.f.martensii.The results showed that the hemolymph exosomes of P.f.martensii are spherical or nearly spherical vesicles with intact membrane structures and clear contours,ranging in size from 75 nm to 150 nm.A total of 101 miRNAs were identified in the PBS and poly(I∶C)groups,of which 76 were known miRNAs.In the poly(I∶C)group,seven miRNAs were significantly upregulated,and five miRNAs were significantly downregulated compared to the PBS group.These differential miRNAs were associated with 570 target genes.KEGG enrichment analysis identified immune-related signaling pathways,including“Acute myeloid leukemia”“JAK-STAT signaling pathway”and“Chemokine signaling pathway”metabolism-related pathways such as“Glycerolipid metabolism”“Purine metabolism”and“Arginine and proline metabolism,”as well as endocrine-related pathways such as“Relaxin signaling pathway”“Prolectin signaling pathway”and“Insect hormone biosynthesis”(P<0.05).Notably,eight miRNAs,including novel_mir2,novel_mir22,miR-2b_5,and miR-279,were found to regulate immune-related signaling pathways and influence the immune response of P.f.martensii by modulating metabolism through FOXO coupling.This study suggests that hemolymph exosomal miRNAs in P.f.martensii coordinately respond to poly(I∶C)stimulation by regulating immune,metabolic,and endocrine processes,providing foundational data for further research into the roles of exosomal miRNAs in the

关 键 词：马氏珠母贝 poly(I∶C) 免疫应答 外泌体 MIRNA 

分 类 号：S917.4[农业科学—水产科学]