白念珠菌粘附素的分离纯化及其在介导与宿主细胞粘附中的作用x.docxVIP

白念珠菌粘附素的分离纯化及其在介导与宿主细胞粘附中的作用白念珠菌粘附素的分离纯化及其在介导与宿主细胞粘附中的作用中华皮肤科杂志 1998年第5期第31卷 论 著作者：刘晓红　廖万清　胡惠民　姚志荣　王君松　赵子卿单位：271000泰安，济南军区第八十八医院皮肤科(刘晓红　王君松　赵子卿)；第二军医大学附属长征医院(廖万清　姚志荣)；第二军医大学生化教研室(胡惠民)关键词：白念珠菌；纤连蛋白；粘附【摘要】　目的　从分子水平探讨白念珠菌(简称白念)与宿主细胞的粘附机制。方法和结果　用非离子去垢剂辛基吡喃葡萄糖苷提取白念孢子壁蛋白质，然后制备纤连蛋白Sepharose 4B亲和柱，运用亲和层析技术从上述提取液中分离纯化纤连蛋白粘附素，在非还原状态进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS)，得到表观分子量60 000和105 000的二条蛋白带。为了弄清此蛋白在白念与宿主细胞粘附中的作用，把人颊粘膜上皮细胞与白念孢子共同振荡孵育2小时，显微镜下测定上皮细胞表面粘附白念的数目，白念与上皮细胞的粘附百分率为44.7±6.28，上皮细胞经纤连蛋白粘附素预处理后，粘附百分率为28.3±6.18，P＜0.01。结论　纤连蛋白粘附素介导了人颊粘膜上皮细胞与白念的粘附。Isolation and Purification of Adhesin of Candida Albican s and its Effect on Adherence to Host CellsLiu Xiaohong, Lia o Wangqing, Hu Huiming, et al. Department of Dermatology, Eighty-eighth Hospita l of Jinan Military Area Command, Taian 271000【Abstract】　Objective　To investigate the adherence mecha nism of Candida albicans to host cells in molecular level.Methods and Results　Yeast cell wall protein of Candida albicans we re extracted, and purified fibronectin (Fn) adhesin was obtained from yeast cell wall extract. Fn adhesin was anal yzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS), the molecular weights of Fn adhesin were 60kd and 105kd. In order to find out t he effect of the protein on adherence to host cells, human buccal epithelial cells and Candida albicans cells we re i ncubated at 37℃ for 2 hours on a rotator, and yeasts adhering to the surfaces o f epithelial cells were assayed microscopically. The adherence rate of C .albicans to buccal epithelial cells was 44.7±6.28%, and after treati n g the epithelial cells with Fn adhesin, the adherence rate reduced to 28.3±6. 18%, P＜0.01.Conclusion　T he results indicate Fn adhesin could mediate adherence of Candida albic ans to epithelial cells.【Key words】　Candida albicans　Fibronectin　Adhere白念珠菌(白念)是重要的条件致病菌，可侵犯皮肤、粘膜和内脏。研究表明白念与宿主细胞间的粘附是其侵入宿主引起感染的前提，在介导白念与宿主细胞的粘附中，纤连蛋白(Fn)起了重要作用［1］。我们运用Fn做亲和配体，用亲和层析技术从白念孢子壁蛋白提取液中成功地分离纯化了Fn粘附素，而且验证了其能介导人颊粘膜上皮细胞与白念的粘附。材料和方法一、材料1.白念为长征医院皮肤科中