胰蛋白酶对烟曲霉分生孢子表面纤连蛋白受体的影响  

The effect of trypsin on surface fibronectin receptors of Aspergillus fumigatus conidia

作　　者：徐赤宇[1] 陈江汉[1] 吴建华[1] 王溪涛[1] 温海[1] 

机构地区：[1]第二军医大学附属长征医院皮肤科,上海200003

出　　处：《临床皮肤科杂志》2005年第8期510-512,共3页Journal of Clinical Dermatology

基　　金：国家自然科学基金资助项目(39770002)

摘　　要：目的:应用流式细胞仪(FCM)测定胰蛋白酶(胰酶)对烟曲霉细胞活力的影响,探讨胰酶对烟曲霉分生孢子表面纤连蛋白受体的影响。方法:用流式细胞仪检测经胰酶预处理20min后的烟曲霉分生孢子的活力、纤连蛋白与烟曲霉分生孢子的黏附率及经胰酶处理后的分生孢子与纤连蛋白黏附率的变化,同时应用激光共聚焦显微镜观察分生孢子表面纤连蛋白受体的分布情况。结果:经胰酶预先处理的分生孢子无凋亡峰出现、被干预的分生孢子接种于马铃薯葡萄糖琼脂(PDA)培养基后仍有大量烟曲霉生长;纤连蛋白与被干预的分生孢子的黏附率随着胰酶浓度的增加而降低;干预后分生孢子表面荧光标记的纤连蛋白受体分布下降。结论:胰酶可降解烟曲霉分生孢子表面相关蛋白受体;流式细胞仪可作为检测烟曲霉分生孢子表面微观变化可靠而高效的辅助手段之一。Objective: To detect the activity of fungal cells pretreated with trypsin by flow cytometry (FCM) and to investi-gate the effects of the trypsin on the fibronectin- receptor of Aspergillus fumigatus conidia. Methods: FCM was used to de-tect the activity of the conidia pretreated with trypsin for 20 min, the binding of fibronectin to the conidia, and adherencerate of the conidia pretreated with trypsin, respectively. Meanwhile we observed the distribution of fibronectin- receptors onthe surface of the conidia by immunofluorescence confocal microscope. Results: There was no apoptosis peak appeared inthe conidia pretreated with trypsin, and dense A. fumigatus colonies grown on PDA with the pretreated conidia were observed.Binding of fibronectin to the conidia was closely dependent and saturable, and the adherence rate of fibronectin and thepretreated conidia decreased with the increase of trypsin concentration. The distribution of FITC labeled Fn- receptors of theconidia pretreated with trypsin was reduced. Conclusions: Trypsin could degrade the protein receptors on the surface of theconidia. FCM not only could be of universal use in cytobiology and in the evaluation of the susceptibility of anti- fungaldrugs, but also could be a highly reliable means to detect the micro changes of the fungal cell surfaces.

关 键 词：烟曲霉 流式细胞仪 胰蛋白酶 

分 类 号：R379.9[医药卫生—病原生物学]