伴放线放线杆菌形态变化对白细胞毒素分泌影响的研究  

Effect of morphological change of Aggregatibacter actinomycetemcomitans on the leukotoxin secretion

作　　者：裴振华[1,2] 王菁[3] 王者玲[1] 

机构地区：[1]首都医科大学口腔医学院牙周粘膜科,北京100050 [2]首都医科大学 [3]北京大学口腔医院第三门诊部

出　　处：《北京口腔医学》2010年第5期253-256,共4页Beijing Journal of Stomatology

基　　金：北京自然科学基金项目(7052029)

摘　　要：目的观察伴放线放线杆菌形态变化对白细胞毒素分泌的影响。方法选择粗糙型和光滑型伴放线放线杆菌各8株,应用聚丙烯酰胺凝胶电泳,检测液体培养12、24、48、60、72h的菌体及培养上清液中116kDa大小白细胞毒素蛋白条带的情况,应用超滤法分离纯化培养上清液蛋白,应用台盼蓝染色排除法检测上清液蛋白白细胞毒素活性。结果粗糙型伴放线放线杆菌菌株液体培养12、24、48、60、72h菌体蛋白电泳均可见116kDa大小的蛋白条带,培养上清液蛋白电泳结果显示116kDa大小的蛋白条带均出现于培养24和48h;光滑型伴放线放线杆菌菌株液体培养12、24、48、60、72h菌体蛋白电泳结果均缺少116kDa大小的蛋白条带,培养上清液蛋白电泳结果显示116kDa大小的蛋白条带出现于培养12和24h;实验菌株培养上清液提取蛋白均具有白细胞毒素活性。结论伴放线放线杆菌粗糙型和光滑型菌株均可分泌具有直接杀灭人多形核白细胞活性的白细胞毒素,但粗糙型菌株分泌白细胞毒素的时间晚于光滑型。Objective To observe the effect of morphological change of Aggregatibacter actinomycetemcomitans on the leukotoxin secretion.Methods SDS-polyacrylamide gel electrophoresis was used to examine leukotoxin（116kDa） production and localization in 8 rough and 8 isogenic smooth stains of A.actinomycetemcomitans after cultured in broth for 12h,24h,48h,60h,72h,and the supernatant protein was purified by ultrafiltration.The activity of secreted leukotoxin in culture supernatants was examined by trypan blue dye exclusion method.Results An intense staining species of 116kDa form cell pellets of different growth ages was found in all 8 rough strains.No 116kDa protein form cell pellets was found in all 8 smooth strains at all time point.The secreted protein had active leukotoxin.Conclusion A.actinomycetemcomitans rough and smooth strains all could secrete active leukotoxin and the rough strains took longer time to produce leukotoxin than smooth strains.

关 键 词：伴放线放线杆菌 形态 白细胞毒素 

分 类 号：R378[医药卫生—病原生物学]