福赛斯坦纳菌体外培养实验技术的研究  

Investigation of Experimental Technique of Tannerella forsythia Cultured in Vitro

作　　者：傅玮涛 苏敏 吴初活 施炜 卢芳国[1] 向琴[2] 谭劲[3] 周小青[2] FU Weitao;SU Min;WU Chuhuo;SHI Wei;LU Fangguo;XIANG Qin;TAN Jin;ZHOU Xiaoqing(Medical School,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Key Laboratory of TCM diagnosis,Hunan University of Chinese Medicine,Changsha,Hunan 410007,China;Stomatology Department,the First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China)

机构地区：[1]湖南中医药大学医学院,湖南长沙410208 [2]湖南中医药大学中医诊断重点实验室,湖南长沙410007 [3]湖南中医药大学第一附属医院口腔科,湖南长沙410007

出　　处：《湖南中医药大学学报》2018年第1期43-46,共4页Journal of Hunan University of Chinese Medicine

基　　金：2016年度湖南省大学生研究性学习和创新性实验计划项目(1021-001017094)

摘　　要：目的探索福赛斯坦纳菌生长繁殖的基本条件,比较N-乙酰胞壁酸及不同营养因子对其生长繁殖的影响,筛选该菌体外培养的最佳营养条件,建立其体外培养的技术方法。方法 (1)培养基制备:在血平板中加0.005%N-乙酰胞壁酸形成改良培养基;制备血平板与不同浓度(0.5、3、10、50、100 mg/L)的N-乙酰胞壁酸形成的培养基;制备10 mg/L N-乙酰胞壁酸与氯化血红素、酵母提取物、维生素K3、脱纤维羊血组合形成不同组别的培养基。(2)接种培养后,用磷酸盐缓冲液将细菌洗脱,并用酶标仪(OD_(600))测定其吸光度。结果福赛斯坦纳菌在含N-乙酰胞壁酸的培养基中生长更好,浓度为10 mg/L时促进作用最明显且细菌形态更为典型。在组3培养基中生长最快。结论 N-乙酰胞壁酸能促进福赛斯坦纳菌的生长繁殖,浓度为10 mg/L时促进作用最明显;N-乙酰胞壁酸及氯化血红素、维生素K3、酵母提取物和脱纤维羊血组合能使其生长周期明显缩短。Objective To investigate the basic requirements for the growth and proliferation of Tannerella forsythia and compare the growth-promoting effects with different nutritional factors and screen the optimum culture in vitro conditions and establish the technique of culturing it.Methods(1)Preparation of culture medium:0.005%N-acetylmuramic acid was added to the blood agar medium,forming the modified medium.Different concentration of N-acetylmuramic acid(0.5,3,10,50,100 mg/L)was added to the blood agar medium.The different mediums of N-acetylmuramic acid(10 mg/L)combined with other nutritional factors(chlorhematin,yeast extract,Vitamin K3,off fiber sheep blood)were formed.(2)After culturing,use PBS to elute the bacterial and measure its absorbance with enzyme-labelled instrument(OD600).Results Tannerella forsythia grew more better in the culture medium containing N-acetylmuramic acid,and the concentration at 10 mg/L was most obvious in growth-promoting effect and can lead to the change of bacterial morphology.The medium which consisted of hemin,yeast extract,menadione and sheep blood was the most suitable condition for its growth.Conclusion N-acetylmuramic acid can promote the growth of Tannerella forsythia,and the combination of N-acetylmuramic acid,hemin,menadione,yeast extract and sheep blood can shorten the growth cycle of it dramatically.

关 键 词：牙周炎 福赛斯坦纳菌 N-乙酰胞壁酸 体外培养 

分 类 号：R965.2[医药卫生—药理学] R780.2[医药卫生—药学]