摘要
为建立大肠埃希菌O_(111)黏附牦牛子宫内膜上皮细胞的致炎模型,体外分离、培养牦牛子宫内膜上皮细胞,培养5d后,加入大肠埃希菌O_(111),通过计算细胞上细菌的黏附数量研究细菌数量和孵育时间对黏附效果的影响,得到最佳黏附条件,通过ELISA法测定细胞培养上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL^(-1)β)的质量浓度变化来鉴定模型。结果表明,大肠埃希菌为1×10~5 CFU/mL、孵育时间为120min是黏附的最佳条件。ELISA法检测结果表明,与对照组相比,60、90、120min组TNF-α、IL-6、IL^(-1)β的质量浓度显著升高,致炎效果明显,表明致炎模型建立成功。
The yak endometrial epithelial cells were isolated and cultured in vitro to establish inflammation models caused by E.coli O111 adhesion yak endometrial epithelial cells.After 5 days＇ culture,Escherichia coli O111 was added in,the optimum condition was obtained by calculation of bacteria number to study the cell adhesion effect under different conditions of bacterial number and incubation time.The optimization model was identified by DAS-ELISA method to determine the content change of tumor necrosis alpha（TNF-α）,leukocyte mediated IL-6 and IL^（-1） factors in cell culture supernatant.The results showed that the number of E.coli was 1×10~5 CFU/mL,and the incubation time was 120 min,which was the best condition for adhesion.ELISA test results proved that the TNF-α,IL-6,IL^（-1）β mass concentration increased significantly and had obvious inflammation effect in 60 min,90 min and 120 min test groups than control group.The model of inflammation was established successfully after series of test.
出处
《西北农业学报》
CAS
CSCD
北大核心
2017年第9期1289-1294,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
西藏自治区自然科学基金(2015212-14-34)
西藏自治区临床兽医学高校重点试验室建设资助项目~~
