摘要
为研究鸭源鸡杆菌对鸡原代输卵管上皮细胞的作用,分别以细菌黏附、侵袭试验及借助姬姆萨染色及扫描电镜观察2株不同来源的鸭源鸡杆菌对输卵管上皮细胞的黏附和侵袭特性。黏附计数表明鸭源鸡杆菌Yu-PDS-RZ-1-SLG能高水平黏附输卵管上皮细胞,而菌株F149T对输卵管上皮细胞有较低的黏附。庆大霉素保护试验结果显示2株菌对输卵管上皮细胞均无可见侵袭力。姬姆萨染色及扫描电镜试验进一步证明输卵管上皮细胞表面有细菌黏附,而内部未见有侵袭的细菌,但Yu-PDS-RZ-1-SLG株接种细胞90min后,细胞逐渐破碎脱落,而菌株F149T则未见明显细胞损伤。菌株经胰蛋白酶处理后,细菌的黏附率显著降低,表明细菌的表面蛋白参与黏附。ELISA法检测鸭源鸡杆菌感染输卵管上皮细胞后炎性细胞因子的变化,结果表明感染组的IL-6、TNF-α、IFN-γ的含量均高于对照组,且Yu-PDS-RZ-1-SLG感染的细胞所分泌的各细胞因子均高于F149T,提示炎性细胞因子的分泌可能改变局部的微环境,促进细菌在输卵管细胞表面增殖,是造成输卵管上皮细胞损伤的机制之一。
To evaluate impact of Gallibacterium anatis （G. anatis ） on primary chicken oviduct epithelial cells （COEC）, adhesion and invasion experiment, Giemsa staining and scanning electron mi- croscopy （SEM） were applied to determine the difference between strains Yu-PDS-RZ-1-SLG and F149T. The results showed that G. anatis strain Yu-PDS-RZ-I-SLG adhered to COEC at a higher level compared to G. anatis F149r and no invasion was observed with gentamicin resistance assay, Giemsa staining and SEM. However, cell debris and cell apoptosis were observed after being exposed to G. anatis Yu-PDS-RZ-1-SLG for 90 rain,while G. anatis F149T did not cause cell damage. Adherence was prevented by treating bacterial cells with trypsin, suggesting the participation of proteins in this process. The levels of cytokines was detected by ELISA after infection,and the resuits showed that IL-6, TNF-α and IFN-γlevels of treatment group were higher than those of control group. Changes of cytokines suggested that the production of cytokines might influence the microenvironment of oviduct, and promote adherence and invasion, which serve as a possible mechanism causing cell damage.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2016年第2期228-233,239,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31302090)
河南省高校科技创新团队支持计划资助项目(14IRTSTHN015)
关键词
鸭源鸡杆菌
原代输卵管上皮细胞
黏附和侵袭
细胞毒性
细胞因子
Gallibacterium anatis
primary chicken oviduct epithelial cells
adherence and invasion
eytotoxicity
cytokines
