新生隐球菌热休克反应对两性霉素B敏感性的影响  

The effect of heat shock reaction in Cryptococcus neoformans on the sensitivity to amphotericin B

作　　者：陈裕充[1] 温海[1] 李志刚[1] 徐红[1] 钟江[2] 尹隽[2] 

机构地区：[1]第二军医大学长征医院皮肤科,上海200003 [2]复旦大学生命科学学院病毒研究室,上海200433

出　　处：《临床皮肤科杂志》2003年第5期254-256,共3页Journal of Clinical Dermatology

摘　　要：目的:探讨新生隐球菌热休克反应在体外药物敏感性实验中对两性霉素B敏感性的影响,以及热休克蛋白70(Hsp70)在新生隐球菌致病中的作用。方法:根据NCCLS的M-27-p方案,测定新生隐球菌在热休克(42℃×30min)前后两性霉素B对新生隐球菌的最小抑菌浓度(MIC)。随机抽取其中7株新生隐球菌,采用鼠抗人、猪、狗等的Hsp70抗体,通过Western印迹法来检测其Hsp70,根据实验信号强弱定性判断在不同刺激条件下的Hsp70产量。结果:①在热休克后两性霉素B对新生隐球菌的MIC较热休克前有明显提高(P=0.002)。②在42℃×30min热应激下新生隐球菌Hsp70产量较在30℃培养下增加。结论:新生隐球菌对两性霉素B的敏感性下降受其热休克反应的影响。结合临床,在较难治愈的病例中Hsp70表达量增加,热休克反应增强;并可能产生与人体Hsp70的交叉免疫反应,推测其对新生隐球菌的致病性可能有一定的作用。Objectives:To inquire the effect of heat shock reaction (HSR)of Cryptococcus neoformans on the sensitivity to amphotericin B (AmB)in vitro and the role of heat shock protein(Hsp70)in the pathogensis of cryptococcosis.Methods:Acc ording to M-27-p protocal,the MIC of AmB in the yeasts was detected before and after heat stress(42℃×30min),respectively.Using the mouse antibody to Hsp70of human,pig,dog,etc,Hsp70was detected from seven randomly sele cted yeast strains by Western blotting.According to the different density of b lotting signal,the production of Hsp70was determined quantatively.Results: ①MIC of AmB increased significantly after stressing in Cryptococci neoformans( P=0.002).②The production of Hsp70in the stressed yeasts was higher th an that cultured in30℃.Conclusions:Decrease in the sensitivity to AmB is re lated to the heat shock reaction(HSR)in Cryptococcus neoformans.The productio n of Hsp70was more in refractory cases with AmB treatment,which indicated the patients infected with the yeasts with strong heat shock reaction were more dif ficult to be cured.The antibody of Hsp70in yeast may have crossing reaction w ith the Hsp70in human.We conferred Hsp70should play a role in pathogenesis of Cryptococcus neoformans.

关 键 词：新生隐球菌 热休克蛋白70 两性霉素B 最小抑菌浓度 WESTERN印迹法 

分 类 号：R379.5[医药卫生—病原生物学]