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罗氏沼虾新型病原体 spiroplasma MR-1008 感染肝胰腺的免疫反应及基因表达

Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen spiroplasma MR-1008.

机构信息

Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China.

出版信息

Fish Shellfish Immunol. 2013 Jan;34(1):315-23. doi: 10.1016/j.fsi.2012.11.009. Epub 2012 Nov 23.

Abstract

Freshwater prawn Macrobrachium rosenbergii inoculated with 100 μl novel pathogen spiroplasma strain MR-1008 in logarithmic phase (10(8) spiroplasmas ml(-1)) were examined for alkaline phosphatase (AKP) activity, acid phosphatase (ACP) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, as well as expressions of 7 immune related genes in hepatopancreas after 1-28 d. Hematoxylin-eosin (HE) staining showed obvious pathological features in hepatopancreas connective and epithelial tissue. Enzyme activity analyze showed that hepatopancreas AKP and ACP activity increased markedly (P < 0.05) when inoculated with spiroplasma MR-1008 after 5 d and 10 d, respectively. SOD enzyme activity changed less obviously and slightly increased at 1 day post-inoculation, but CAT activity decreased significantly after 5 d inoculation. The expression levels of lipopolysaccharide and β-1,3-glucan-binding protein (LGBP), peroxinectin (PE), α2-macroglobulin (α2M), AKP, ACP, CAT, and copper/zinc SOD (Cu, Zn-SOD) genes in the hepatopancreas were examined by Real-Time PCR (qRT-PCR) and the results demonstrated that these immune related genes were induced by challenge with spiroplasma MR-1008. The results suggested that the prawn immune responses could be activated or inhibited by spiroplasma MR-1008, and that the hepatopancreas also plays key roles in innate immunity for defense against the pathogen.

摘要

罗氏沼虾(Macrobrachium rosenbergii)经对数生长期(10(8) 螺旋体 ml(-1))的新型病原体螺旋体菌株 MR-1008 接种 100 μl 后,连续 1-28 天检测其肝胰腺碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及 7 种免疫相关基因的表达情况。肝胰腺组织苏木精-伊红(HE)染色显示明显的病理特征。酶活性分析表明,接种螺旋体 MR-1008 后 5 天和 10 天,肝胰腺 AKP 和 ACP 活性显著增加(P < 0.05)。SOD 酶活性变化不明显,接种后 1 天略有增加,但 5 天接种后 CAT 活性显著下降。采用 Real-Time PCR(qRT-PCR)检测肝胰腺中脂多糖和β-1,3-葡聚糖结合蛋白(LGBP)、过氧化物酶(PE)、α2-巨球蛋白(α2M)、AKP、ACP、CAT 和铜/锌 SOD(Cu,Zn-SOD)基因的表达水平。结果表明,这些免疫相关基因被螺旋体 MR-1008 诱导。结果表明,罗氏沼虾的免疫反应可能被螺旋体 MR-1008 激活或抑制,肝胰腺在先天免疫防御病原体中也发挥关键作用。

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