【摘要】：甘露糖受體(MR)隸屬凝集素超家族,主要表達于巨噬細胞和未成熟的樹突狀細胞表面。MR不僅在先天免疫防御中發(fā)揮重要作用,還通過參與抗原呈遞,激活T淋巴細胞,啟動獲得性免疫應答過程。本研究采用同源克隆技術獲得黃顙魚甘露糖受體(pf MR)基因,采用熒光定量PCR技術檢測MR在正常黃顙魚體內的分布情況,采用湒愛德華菌感染黃顙魚頭腎巨噬細胞和甘露聚糖封閉MR方法研究黃顙魚MR在抗細菌感染中的作用。結果顯示,pf MR同團頭魴、草魚、斑馬魚和尼羅羅非魚的MR聚為一支。pf MR在所檢測的12個組織中均有分布,其在腎臟、脾臟和肌肉組織中表達量較高,在血液中表達量較少。湒愛德華菌感染黃顙魚頭腎巨噬細胞后,pf MR、IL-1β和TNF-a均被細菌誘導表達,超氧陰離子和一氧化氮的含量也上升,超氧陰離子在感染30 min后即顯著上升,一氧化氮在感染12 h后才顯著上升。甘露聚糖競爭結合MR,顯著抑制巨噬細胞內化GFP標簽的湒愛德華菌,加入EDTA減少內化的熒光強度,加入Ca~(2+)使內化的熒光強度回升。研究表明,黃顙魚頭腎巨噬細胞MR參與湒愛德華菌的識別和內吞過程,而且依賴Ca~(2+)。
[Abstract]:Mannose receptor (MR) belongs to the lectin superfamily and is mainly expressed on macrophages and immature dendritic cells. Mr not only plays an important role in innate immune defense, but also activates T lymphocytes by participating in antigen presentation. Initiate the process of acquired immune response. In this study, the mannose receptor (pf MR) gene of Pelteobagrus fulvidraco was obtained by homologous cloning technique, and the distribution of MR in normal yellow catfish was detected by fluorescence quantitative PCR. The role of MR in anti-bacterial infection of Pelteobagrus fulvidraco MR was studied by using Edwardian bacteria infection and mannan blocking MR method in the head kidney macrophages of Pelteobagrus fulvidagrus. The results showed that the MR of, pf MR was the same as that of snout bream, grass carp, zebrafish and Nile tilapia. The expression of pf MR was higher in kidney, spleen and muscle than in blood. The expression of pf MR,IL-1 尾 and TNF-a were induced by bacteria, the contents of superoxide anion and nitric oxide increased, and the superoxide anion increased significantly at 30 min after infection with Evodomonas fulvidracobagrus, the expression of SOD 尾 and TNF-a were induced by bacteria, and the content of superoxide anion and nitric oxide increased significantly after 30 min infection. No increased significantly after 12 hours of infection. The competition of mannan combined with MR, significantly inhibited the internalization of GFP-labeled Eduardo spp. by adding EDTA, decreased the fluorescence intensity of internalization, and increased the fluorescence intensity of internalization by adding Ca~ (2). The results showed that MR was involved in the identification and endocytosis of Edwards and depended on Ca~ (2).
【作者單位】： 華中農業(yè)大學水產學院水生動物醫(yī)學系;華中農業(yè)大學水產學院淡水水產健康養(yǎng)殖湖北省協同創(chuàng)新中心;華中農業(yè)大學水產學院池塘健康養(yǎng)殖湖北省工程實驗室;華中農業(yè)大學水產學院農業(yè)部淡水生物繁育重點實驗室;
【基金】：國家自然科學基金(31372563,31572657,31502199) 中央高�；究蒲袠I(yè)務費專項(2014PY035,2662015QC019) 湖北省科技支撐計劃(2015BBA228,YSF2015000255) 農業(yè)部漁用藥物創(chuàng)制重點實驗室開放課題(201401) 廣東省海洋與漁業(yè)局基金(A201512C003,2015-115) 武漢市科技局基金(2016020101010089)~~
【分類號】：S943

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