创伤弧菌溶细胞素融合蛋白对小鼠单核巨噬细胞产NO及其合酶的影响  

Study on expression of NO and iNOS in J774A. 1 cells induced by rVvhA

作　　者：杜李梅[1] 郭雅君[1] 屠艳烨[1] 胡蝶[1] 楼永良[1] 严杰[2] 

机构地区：[1]温州医学院检验医学院,浙江省医学遗传学重点实验室,325035 [2]浙江大学医学院病原生物学教研室

出　　处：《中华微生物学和免疫学杂志》2010年第10期887-891,共5页Chinese Journal of Microbiology and Immunology

基　　金：基金项目：浙江省科技计划（2008C33058）;浙江省自然科学资金（Y2090468）

摘　　要：目的 研究创伤弧菌溶细胞素融合蛋白（rVvhA）对小鼠单核-巨噬细胞（J774A.1）产生一氧化氮（NO）和诱导型一氧化氮合酶（iNOS）表达的影响.方法 应用MTT法检测rVvhA对J774A.1增殖的抑制作用;Griess法检测NO含量;RT-PCR和免疫荧光法检测iNOS mRNA和蛋白表达水平.结果 0.8 HU/ml及以上浓度rVvhA可显著抑制J774A.1的增殖;在IFN-γ的辅助作用下,0.4 HU/ml的rVvhA即可诱导J774A.1产生大量NO,显著增加细胞内活性以及iNOS mRNA和蛋白表达水平.结论 rVvhA可诱导巨噬细胞产生NO和iNOS表达,在创伤弧菌的致病机制中具有重要意义.Objective To investigate the effect of recombinant Vibrio vulnificus hemolysin （rVvhA） on the expression of nitric oxide（NO） and induced nitric oxide synthase（iNOS） in J774A. 1 cells.Methods The inhibitory effect of rVvhA on J774A. 1 proliferation was measured by MTF colorimetry technique. The content of nitrite in culture medium was determined by Griess reagent. The expression of iNOS protein and mRNA were measured by immunofluorescence and RT-PCR, respectively. Results The viability of J774A. 1 cells was apparently inhibited when exposed to 0.8 HU/ml rVvhA and up. With the help of IFN-γ, the expression of NO and the activity of iNOS in J774A. 1 cells were remarkably increased when exposed to 0.4 HU/ml rVvhA. Conclusion rVvhA can increase the expression of NO and iNOS, it may play an important role in the pathogenic mechanism of Vibrio vulnificus.

关 键 词：创伤弧菌 溶细胞素 J774A.1 一氧化氮 诱导型一氧化氮合酶 

分 类 号：R743.31[医药卫生—神经病学与精神病学]