创伤弧菌攻击对人脐静脉内皮细胞系的影响

Influence of Vibrio vulnificus attack on human umbilical vein endothelial cell line

背景:创伤弧菌感染后患者死亡率高达50%以上,且感染后大量创伤弧菌侵入血液之中,而血管内皮细胞是阻挡创伤弧菌进入血管的第一道屏障。目的:观察创伤弧菌对人脐静脉内皮细胞增殖能力和细胞形态的影响。方法:将对数生长期的人脐静脉内皮细胞消化后,调整细胞浓度至5×108cfu/L,依培养基中创伤弧菌细菌浓度不同分为5个组:104,105,106,107cfu/L创伤弧菌组和对照组。对照组不加创伤弧菌,仅用体积分数为10%胎牛血清DMEM培养基培养。锥虫蓝拒染法检测细胞存活率、MTT法观察创伤弧菌对人脐静脉内皮细胞增殖的影响,及光镜下观测细胞面积、周长、细胞核的面积、核浆比以及形状不规则指数。结果与结论:随着创伤弧菌作用浓度和时间的增加,人脐静脉内皮细胞的增殖能力逐渐减弱,攻击组细胞的面积和周长,细胞核的面积以及核浆比都比对照组明显增大,形状不规则指数比对照组更偏离。提示创伤弧菌对体外培养的人脐静脉内皮细胞系的增殖有明显的抑制作用,其抑制能力与创伤弧菌作用的浓度和时间有关。

BACKGROUND：Mortality of Vibrio vulnificus infection is more than 50% in patients,and a large amount of Vibrio vulnificus intrude into the blood.Vascular endothelial cells are the first barrier to stop Vibrio vulnificus.OBJECTIVE：To observe effect of Vibrio vulnificus on proliferation and morphology of human umbilical vein endothelial cells.METHODS：The human umbilical vein endothelial cells at log growth phase were digested,adjusted at a density of 5×10^8 cfu/L and incubated with Vibrio vulnificus（10^4,10^5,10^6,10^7cfu/L）.In addition,cells cultured with DMEM medium containing 10% fetal bovine serum served as normal control group.Using trypan blue exclusion staining method was used to detect cell survival;MTT method to observe influence of Vibrio vulnificus on cell proliferation;cell area and circumference,nucleus area,ratio of nucleus and cytoplasm,as well as irregular shape index were observed by light microscopy.RESULTS AND CONCLUSION：With increasing concentration and duration of Vibrio vulnificus,cell proliferation was gradually weakened.The cell area and circumference,the nucleus area,and the ratio of nucleus and cytoplasm were significantly increased compared with the control group;the irregular shape index was more deviated than the control group.Results show that Vibrio vulnificus significantly inhibits proliferation of cultured human umbilical vein endothelial cells,and the inhibition relates to the concentration and duration of Vibrio vulnificus.