巨噬细胞与三株念珠菌作用后MyD88和CARD9基因表达的研究

目的：通过检测三株不同白色念珠菌作用下，巨噬细胞激活的关键下游信号传导分子CARD9、MyD88基因表达水平以及所产生的活性氧ROS差异，探讨巨噬细胞对三株白色念珠菌杀伤能力产生差异的可能原因。方法：将骨髓来源小鼠巨噬细胞分别与三株念珠菌以l：10的比例共培养，30min、1h、2h和4h收集巨噬细胞，荧光定量PCR检测MyD88、CARD9基因的表达差异，采用流式细胞术检测所产生的ROS量。结果：RT－PCR结果显示，巨噬细胞与三株白色念珠菌共培养早期MyD88基因表达水平，3683组最高；CARD9基因表达水平，3630组最高；随着时间的增长，不同组别的MyD88基因相对表达无明显变化，而CARD9基因共培养2h相对表达量SC5314组高于3630组及3683组（P〈0．01）；4h，3683组相对表达量高于3630组、SC5314组（P〈0．01）。流式细胞检测结果显示ROS荧光强度3630组〉3683组〉SC5314组（p〈o．05）。结论：CARD9是白色念珠菌PAMPs与PRRs识别及产生抗念珠菌感染关键分子ROS信号通路的关键分子，其表达水平的差异有可能是导致巨噬细胞对不同念珠菌杀伤能力产生差异的机制之一。’

Empression of MyD88 and CARD9 in Responses of Murine Macrophage t0~ Three Strains of Candida Albicans.

Objective： To investigate ; the gene expression of MyD88 and CARD9 in responses of murine macro phage to three strains of Candida albicans. Methods： Macrophages and three strains of C. albicans were cocultured at a ratio of 1：10. Cell mixtures were collected at time points of 30min, lh, 2h, 4h and stored at -80℃for expres- ＇! sion of MyD88, CARD9 genes with fluorescent quantitative RT-PCR. Results： After macrophages cocultured with three strains of C. albicans, in ea？ly time, MyD88 gene expression of macrophages induced by strain 3683 and CARD9 gene expression induced by strain 3630 were the hightest when comparing with those induced by the other strains of yeasts. The gene expression level of MyD88 did not change at lh, 2h and 4h time points. However, 2h after coculture with sC5314, the gene expression of CARD9 was higher than that coculture with 3630 and 3683 （P %0.01）; and at 4h , the gene expression of CARD9 in 3683 coculture group was higher than that in 3630 and SC5314 group （P〈0.01）. Conclusion： CARD9 could be seen as a key factor of the signaling pathway in activation of macrophages and which may lead to the varieties of maerophages killing of different strains of yeasts.