布鲁菌S2株感染脑微血管内皮细胞体外模型的构建

布鲁菌S2株感染脑微血管内皮细胞体外模型的构建
徐婷;李海宁;张吉;杨娟;王国玮;王振海
【期刊名称】《宁夏医科大学学报》
【年(卷),期】2018(040)002
【摘要】Objective Construction of Brucella S2 infected brain microvascular endothelial cells in vitro model. Methods Brucella S2 strain was cultured in soybean casein medium for 4 days. Gram staining and Kirzovsky staining were used for bacterial identification. Brucella S2 strain infected brain microvascular endothelial cells, and the intracellular status of Brucella was observed by transmission electron microscopy and intracellular colony counts. observed the effect of Brucella S2 strain on cell membrane by cell viability and lactate dehydrogenase activity,peripheral blank contral group. Flow cytometry was used to observe the apoptosis of brain microvascular endothelial cells infected by Brucella S2 strain,peripheral blank control group. Results Transmission electron microscopy showed that the S2 strain of Brucella could infect brain microvascular endothelial cells and exist in endocytic vacuoles. Intracellular colony counting method was used to count intracellular bacteria. The amount of intracellular bacteria increased with the prolongation of transfection time.The bacterial population showed an increasing trend, and there was a significant difference between the intracellular bacteria at each time point(P<0.01). Trypan blue staining results showed that the blank group and the treatment group how
to illustrate the group. There was no significant difference in the cell survival rate between the two groups(P>0.05). In the early stage of infection(before 12h),the activity of lactate dehydrogenase in cells increases and the cell membrane were damaged. At 24h to 48h,the intracellular activity of lactate dehydrogenase was low,and the damage to the cell membrane is small. Flow cytometry showed no significant difference in apoptosis rate in blank group or treatment group(P>0.05). Conclusion This experiment is successfully constructed an in vitro model of Brucella S2 strain infecting brain microvascular endothelial cells,and confirmed that Brucella S2 strain can enter the brain microvascular endothelial cells and survive intracellularly.%目的构建布鲁菌S2株感染脑微血管内皮细胞的体外模型.方法布鲁菌S2株采用大豆酪蛋白培养基培养4d,通过革兰染色及柯兹罗夫斯基染色进行细菌鉴定;布鲁菌S2株感染脑微血管内皮细胞后,通过透射电镜和胞内菌落计数观察布鲁菌进入胞内的情况;通过细胞存活率和乳酸脱氢酶活性观察布鲁菌S2株对细胞膜的影响,外设空白对照组;采用流式细胞仪观察布鲁菌S2株感染脑微血管内皮细胞凋亡的情况,外设空白对照组.结果透射电镜显示布鲁菌S2株能够感染脑微血管内皮细胞并存在于内吞泡内,胞内菌落计数法统计细胞内菌量发现,胞内活菌量随转染时间的延长而增加(P<0.05),感染48h胞内菌落数高于12h和24h(P<0.05),感染12h和24h胞内菌落数差异无统计学意义(P>0.05);台盼蓝染色结果显示空白组与处理组间细胞存活率差异无统计学意义(P>0.05);在感染初期(12h前)细胞内的乳酸脱氢酶活性会增高,会对细胞膜造成损伤,在24h到48h的时候,细胞内的乳酸脱氢酶活活性较低,对细胞膜的损伤较小;流式细胞仪检测显示空白组与处理组间细胞凋亡率差异无统计学意义(P>0.05).结论
本实验成功构建布鲁菌S2株感染脑微血管内皮细胞体外模型,明确了布鲁菌S2株能够进入脑微血管内皮细胞并在胞内存活.
【总页数】5页(P181-185)
【作者】徐婷;李海宁;张吉;杨娟;王国玮;王振海
【作者单位】宁夏医科大学,银川750004;宁夏颅脑疾病重点实验室,银川750004;宁夏医科大学总医院神经病学中心,银川750004;宁夏医科大学,银川750004;宁夏颅脑疾病重点实验室,银川750004;宁夏医科大学总医院神经病学中心,银川750004;宁夏医科大学,银川750004;宁夏颅脑疾病重点实验室,银川750004;宁夏颅脑疾病重点实验室,银川750004;宁夏医科大学总医院神经病学中心,银川750004
【正文语种】中文
【中图分类】R74
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