羊种布鲁杆菌外膜蛋白31诱导BV-2小胶质细胞自噬体形成  

Outer membrane protein 31 of B. melitensis induces the formation of autophagosomes in BV-2 microglia cells

作　　者：王钊[1] 王妍柏[2] 张吉[1] 徐婷[1] 林涛[1] 马博雅[1] 王振海[2,3] 

机构地区：[1]宁夏医科大学 [2]宁夏医科大学总医院 [3]宁夏颅脑疾病重点实验室,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2017年第1期17-21,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81260189)

摘　　要：目的确定羊种布鲁杆菌外膜蛋白31(Omp31)对BV-2小胶质细胞自噬形成的影响。方法 (0.17、0.50、1.50、4.50、13.50)μg/m L Omp31分别处理BV-2细胞6 h,实时定量PCR检测微管相关蛋白1轻链3B(LC3B)mRNA的表达,Western blot法检测LC3BⅡ蛋白、LC3B蛋白的表达;采用透射电镜技术观察各组细胞自噬体,ELISA检测培养细胞上清液肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10的水平。结果 Omp31处理增加BV-2小胶质细胞LC3BⅡ蛋白水平,且0.5μg/m L Omp31处理LC3BⅡ蛋白增加最明显;0.5μg/m L以下的Omp31对LC3B mRNA的表达有促进作用,大于0.5μg/m L Omp31则抑制LC3B mRNA的表达;0.5μg/m L Omp31能诱导BV-2细胞形成较多自噬体;Omp31能诱导BV-2细胞表达TNF-α、IL-6,但抑制IL-10的表达。结论 0.5μg/m L剂量的Omp31能诱导BV-2细胞发生明显自噬。Objective To determine the influence of outer membrane protein 31（ Omp31） of B. melitensis on the formation of autophagosomes in BV-2 microglia cells. Methods BV-2 cells were treated for 6 hours with Omp31 at 0. 17,0. 50,1. 50,4. 50,13. 50 μg/m L. Real-time quantitative PCR was used to detect the expression of microtubule associated protein 1 light chain 3B（ LC3B） mRNA; Western blotting was applied to detect the expressions of LC3 B Ⅱ and LC3 B proteins; transmission electron microscopy was performed to observe cell autophagy in each group. The levels of tumor necrosis factor α（ TNF-α）,interleukin 6（ IL-6） and IL-10 in the culture cell supernatant were determined with ELISA. Results Omp31 treatment resulted in an increase in the level of LC3 B Ⅱ protein,especially the most obvious increase in the ones treated by 0. 5 μg/m L Omp31.The concentration of Omp31,being no more than 0. 5 μg/m L,could promote the expression of LC3 B mRNA,while more than0. 5 μg/m L Omp31,it could inhibit the expression of LC3 B mRNA; 0. 5 μg/m L Omp31 could induce the formation of more autophagosomes in BV-2 cells. Omp31 promoted the expressions of TNF-α and IL-6,and inhibited the expression of IL-10 in BV-2 cells. Conclusion Omp31 at the concentration of 0. 5 μg/m L can significantly induce autophagy in BV-2 cells.

关 键 词：羊布鲁杆菌 外膜蛋白 小胶质细胞 自噬 炎症 

分 类 号：R392.12[医药卫生—免疫学]