目的研究新生隐球菌体外对角质形成细胞活力的影响。方法将新生隐球菌父代标准株与子代荚膜缺陷株于体外分别与角质形成细胞分别共培养,同时设立热灭活的菌体、空白对照,再分别设立菌体与细胞直接接触与不接触共培养相互对照,分别作用0.5 h、1 h和2 h后,采用流式细胞仪检测隐球菌角质形成细胞的调亡率。结果随着时间延长,与空白对照组及热灭活组比较,实验组角质形成细胞的凋亡率逐渐增加。无荚膜株与父代有荚膜株比较,无荚膜株对细胞活力的影响在作用后1 h、2 h明显低于有荚膜株。2种菌株不直接接触培养使细胞的凋亡率明显下降;不直接接触的有荚膜株与热灭活的菌体之间比较差异不显著。结论虽然有荚膜株与无荚膜株隐球菌均可以使角质形成细胞活性明显降低,但荚膜可以显著增强菌体对细胞活力的影响;角质形成细胞活力的降低主要是通过与菌体接触培养后产生的,诱导细胞调亡需要菌体与细胞的直接接触。
Objective To investigate the effect of Cryptococcus neoformans on the viability of keratinocytes.Methods Strains of Cryptococcus neoformans B3501 and its acapsule mutant cap64 were cocultured with keratinocytes and the viability of the keratinocytes was measured by flow cytometry after 0.5 h,1 h and 2 h.Results were compared with those of negative control,hear-killed fungi,and the fungi indirectly contacted with keratinocytes.Results Viability of keratinocytes decreased significantly cocultured with Cryptococcus neoformans by time prolonging,while more significantly after 1 h and 2 h's coculture with B3501.There was no difference in the viability of keratinocytes between coculture with killed B3501 and indirect coculture with B3501.Conclusions Capsule of Cryptococcus neoformans plays an important role in viability of keratinocytes.Indirect contact with Cryptococcus neoformans is necessary for viability decreasing.