摘要
为检测磷脂酰肌醇3激酶(PI3K)抑制剂LY294002对单核细胞增生李斯特菌(Listeria monocytogenes,L.monocytogenes)在小鼠脑微血管内皮细胞内存活的影响。将不同浓度抑制剂LY294002与细胞共同孵育,MTT法检测不同浓度抑制剂LY294002对细胞活性的影响,然后将L.monocytogenes分别感染细胞,利用菌落计数法计算L.monocytogenes的胞内细菌数量,检测LY294002对L.monocytogenes介导的小鼠生存曲线的影响,利用试剂盒检测抑制剂LY294002对L.monocytogenes介导的LDH和IFN-γ分泌的影响。抑制剂LY294002在5μmol/L、10μmol/L或20μmol/L浓度时不影响细胞活性,LY294002显著抑制了L.monocytogenes的胞内和脏器内细菌数量,提高L.monocytogenes介导的小鼠存活率,分别显著提高和降低L.monocytogenes介导的IFN-γ水平和LDH水平(P<0.05)。本研究表明,LY294002可调控细胞内Lm的存活,为L.monocytogenes引起脑膜炎的致病机制提供科学依据。
To examine the effect of LY294002,a phosphatidylinositol 3-kinase（ PI3 K） inhibitor,on the survival of（ Listeria monocytogenes,L. monocytogenes） in mouse brain microvascular endothelial cells,different concentrations of inhibitor LY294002 were co-incubated with cells. The effect of different concentrations of inhibitor LY294002 on cell viability was measured by MTT assay.L. monocytogenes was then inoculated to cells and the number of intracellular bacteria was calculated using colony counting method.The influence of the survival curve of the mice was examined using the kit to examine the effect of inhibitor LY294002 on L. monocytogenes-mediated secretion of LDH and IFN-γ. The inhibitor LY294002 did not affect the cell activity at concentrations of 5μmol/L,10 μmol/L,or 20 μmol/L. LY294002 significantly inhibited the intracellular and intracellular bacterial counts of L. monocytogenes and increased the survival rate of mice mediated by Lm. Significantly increased and decreased L. monocytogenes-mediated IFN-γ levels and LDH levels（ P〈 0. 05） were observed,respectively. Our data shew that LY294002 can regulate the survival of intracellular L. monocytogenes.
作者
张俊波
印双红
易萌
张红
ZHANG Jun-bo;YIN Shuang-hong;YI Meng;ZHANG Hong(Cultural and Technological Industry Innovation Research Center）Tongren University,Tongren 554300,Chin;College of One Health,Tongren University,Tongren 554300,China;College of Big Data Institute,Tongren University,Tongren 554300,China])
出处
《中国兽医杂志》
CAS
北大核心
2018年第5期14-18,共5页
Chinese Journal of Veterinary Medicine
基金
国家自然科学基金项目(31502067)
中国博士后科学基金资助项目(2017M613003)
铜仁学院博士科研启动基金(trxy DH1504)
贵州省教育厅自然科学研究重点项目[黔教合KY字(2015)411号]
贵州省教育厅青年科技人才成长项目[黔科合KY字(2017)312]
贵州省科技合作计划项目[黔科合LH字(2015)]7238
黔科合LH字(2015)7236号]
