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免疫调节基因精氨酸酶-2和诱导型一氧化氮合酶(iNOS)在两种虹鳟(Oncorhynchus mykiss)品系暴露于粘孢子虫后表达的研究。

Expression of immune-regulatory genes, arginase-2 and inducible nitric oxide synthase (iNOS), in two rainbow trout (Oncorhynchus mykiss) strains following exposure to Myxobolus cerebralis.

机构信息

Clinic for Fish and Reptiles, Faculty of Veterinary Medicine, University of Munich, Kaulbachstrasse 37, 80539 Munich, Germany.

出版信息

Parasitol Res. 2010 Jan;106(2):325-34. doi: 10.1007/s00436-009-1661-0. Epub 2009 Nov 6.

Abstract

The present endeavor was conducted to evaluate the role of activated macrophage in the susceptibility of two different rainbow trout (Oncorhynchus mykiss) strains, a susceptible American (T) and a more resistant German (H), to infection with Myxobolus cerebralis. Arginase-2 and inducible nitric oxide synthase (iNOS) genes were used as references to the alternative and classical pathway of macrophage activation. The expression level of both genes was measured using quantitative real-time polymerase chain reaction. The expression level of arginase-2 was significantly upregulated in strain T at 2 h and 8 days post exposure in the strain H. In case of iNOS, the expression level was significantly upregulated from 24 h to 8 days p.e. in strain T and only in 8 days p.e. in strain H. During this study also, the influence of nitric oxide (NO) on the viability of the triactinomyxon spores (TAMs) of M. cerebralis was evaluated using the NO-donor S-nitroso-N-acetyl-penicillamine (SNAP). Rising final concentrations of SNAP from 0.25 to 1 mM at 2, 4, and 24 h resulted in increasing numbers of propidium iodide-positive TAMs detected. The results of this study suggest an inability of strain T to react with an effective immune response against infection with M. cerebralis. Furthermore, the TAMs of M. cerebralis react with significant decrease of viable spores to rising concentration of SNAP and longer incubation, but there is also evidence for some resistance to NO activity.

摘要

本研究旨在评估激活的巨噬细胞在两种不同虹鳟鱼(Oncorhynchus mykiss)品系(易感的美国品系(T)和更具抗性的德国品系(H))对粘孢子虫(Myxobolus cerebralis)感染易感性中的作用。精氨酸酶-2 和诱导型一氧化氮合酶(iNOS)基因被用作巨噬细胞激活的替代和经典途径的参考。使用定量实时聚合酶链反应测量这两个基因的表达水平。在暴露后 2 小时和 8 天,T 品系中精氨酸酶-2 的表达水平显著上调;而在 H 品系中,从 24 小时到 8 天 p.e.,iNOS 的表达水平显著上调。在本研究中,还使用一氧化氮(NO)供体 S-亚硝基-N-乙酰青霉胺(SNAP)评估了 NO 对粘孢子虫(M. cerebralis)三射虫孢子(TAMs)活力的影响。随着 SNAP 终浓度从 0.25 到 1 mM 的升高,在 2、4 和 24 小时检测到的碘化丙啶阳性 TAMs 的数量逐渐增加。本研究结果表明 T 品系无法对粘孢子虫(M. cerebralis)的感染产生有效的免疫反应。此外,M. cerebralis 的 TAMs 对 SNAP 浓度升高和孵育时间延长的反应导致存活孢子数量显著减少,但也有证据表明对 NO 活性有一定的抗性。

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