Eriocheir sinensis vesicle-associated membrane protein can enhance host cell phagocytosis to resist Spiroplasma eriocheiris infection
Eriocheir sinensis vesicle-associated membrane protein can enhance host cell phagocytosis to resist Spiroplasma eriocheiris infection
复制标题
中华绒螯蟹囊泡相关膜蛋白可增强宿主细胞的吞噬能力，抵抗绒毛螺原体感染。
DOI：
10.1016/j.fsi.2022.08.012
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发表时间：
2022-08-26
影响因子：
4.7
通讯作者：
Meng，Qingguo
中科院分区：
文献类型：
--
作者：
Wang，Yaqin;Miao，Yanyang;Meng，Qingguo
囊泡相关膜蛋白（VAMP）是分泌性转运囊泡膜上的受体蛋白，参与宿主免疫功能。胞内病原体绒螯螺原体可引起中华绒螯蟹震颤病。在以前的一项研究中，它被发现。蛋白质组学分析表明，中华绒螯蟹VAMP（EsVAMP）在感染绒螯蟹后有不同程度的表达。本研究旨在探讨EsVAMP在S.绒螯菌感染EsVAMP基因全长1681 bp，包含一个395 bp的开放阅读框，90 bp的5′-非编码区和1277 bp的3′-非编码区。qPCR结果表明，EsVAMP在血细胞和神经中表达量最高，其次是鳃、肠和肝胰腺，在心脏和肌肉中表达量较低。绒螯菌感染EsVAMP过表达后，S.实验组RAW 264. 7细胞形态和细胞活力明显优于对照组。同时，S.实验组羊毛蕨素显著低于对照组。将EsVAMP与pCMV-Cre-mCherry连接后转染RAW264.7细胞，发现EsVAMP与溶酶体共定位。同时，吞噬的S.巨噬细胞吞噬率和巨噬细胞吞噬率均显著提高。通过合成EsVAMPdsRNA进行干扰实验，以验证EsVAMPmRNA被成功抑制。帖前EsVAMPRNAi和S.绒螯菌感染同时，吞噬的S. EsVAMPRNAi和S.绒螯菌感染这些结果表明VAMP参与细胞吞噬作用以抵抗病原体感染。
Vesicle-associated membrane protein (VAMP) belongs to the receptor protein on the membrane of the secretory transport vesicle and involves in host immune function. The intracellular pathogenSpiroplasma eriocheiriscould causeEriocheir sinensistremor disease. In a previous study, it was foundE. sinensisVAMP (EsVAMP) was differently expressed inS. eriocheirisinfection by proteomics analysis. This study mainly aims at the function of EsVAMP in the process of theS. eriocheirisinfection. The length ofEsVAMPgene was 1681 bp, which contained a 395 bp open reading frame, 90 bp 5′-non-coding region (UTR) and 1277 bp 3′-UTR. The results of qPCR showed thatEsVAMPwas expressed highly in hemocytes and nerves, followed by gills, intestines and hepatopancreas, and lowly expressed in heart and muscles.EsVAMPin hemocytes was up-regulated afterS. eriocheirisinfection. After EsVAMP over-expression andS. eriocheirisinfection, the RAW264.7 cell morphology and cell viability of the experiment group were significantly better than the control group. Meanwhile, the copy number ofS. eriocheirisin the experiment group was significantly lower than that in the control group. After EsVAMP and pCMV-Cre-mCherry were ligated and transfected into RAW264.7 cells, it was found that EsVAMP and lysosome co-localized. Meanwhile, the phagocytosed inactivatedS. eriocheirisnumber and phagocytosed efficiency in RAW264.7 cells were increased significantly. The interference experiment was carried out by synthesizingEsVAMPdsRNA to verify that theEsVAMPtranscriptions were successfully suppressed. TheS. eriocheiriscopy number and the mortality of crab increased significantly afterEsVAMPRNAi andS. eriocheirisinfection. Meanwhile, the phagocytosed inactivatedS. eriocheirisnumber and phagocytosed efficiency in hemocytes decreased significantly afterEsVAMPRNAi andS. eriocheirisinfection. These results showed that VAMP was involved in the cell phagocytosis to resist pathogen infection.