Oligo基因芯片和小鼠模型研究外阴阴道念珠菌病的发病机制

目的 探讨外阴阴道念珠菌病的发病机制。方法 将来自感染组、阴性对照组及空白对照组小鼠模型的标本分别与自制的疾病相关因子基因表达谱芯片杂交，将感染组与对照组中各疾病相关因子信号值进行比较，筛选出表达升高或降低2倍的因子，并绘制成差异基因表达谱进行分析。结果 与空白对照组相比在感染组中可以出现表达上调的基因有39个，可以出现表达下调的基因有4个。在宿主免疫方面，炎性趋化因子普遍表达增高，适应性免疫调节因子IL-1、IL-4、IL-6、IL-10、IL-12、TNF-α、NF-κB以及TGF-β均有不同程度增强，全部标本天然免疫成分TLR4的表达增高，TLR2仅表达增高于1/3的标本。在致病菌株毒力方面，菌丝生成调控因子EFG1、分泌型天冬氨酸蛋白酶（SAP）2、4、5、6、10、脂酶（LIP）2、LIP4和菌丝胞壁蛋白（HWP）1表达显著增强。其中，单核细胞趋化蛋白（MCP）-1、巨噬细胞炎性蛋白（MIP）-1α和MIP-2、IL-1、Toll样受体（TLR）4、LIP4、HWP1在全部感染组标本中显著增高，这些因子涉及白念珠菌胞外水解酶、菌丝形成、表型转换及宿主天然免疫过程。结论 适应性免疫的功能不足以及致病菌株的毒力增强均参与小鼠阴道念珠菌病的发病机制，TLR4在该病局部宿主免疫机制中可能起较为重要的作用。

Pathogenesis analysis of vulvovaginal candidiasis in a mouse model using oligonucleotide microarrays

Objective To investigate the pathogenesis of vulvovaginal candidiasis.Methods Thirty mice were randomly and equally divided into 3 groups,i.e.,infected group treated with estrogen and innoculated with a clinical isolate of C. albicans,negative centrel group treated with estrogen and inoculated with physiological saline.blank control group without any treatment.All mice were kilied on day 7 after inoculation,vaginal tissue samples were obtained and subjected to pathological examination and staining.Total RNA was extracted from these samples as well as Candida isolates and reference strains,and hybridized with self-designed oligonucleotide chips.Then,signal value of vulvovaginal candidiasis-associated factors was compared between infected group and blank control group,and a 2-fold difference was considered as significant.Results Compared with the blank centrol group,the gene expression of 39 factors increased while that of 4 factors decreased in the infeeted group.In the case of host immunity,the expression of inflammatory chemokines generally inereased,and that of regulatory factors of adaptive immunity,including interleukin (IL)-1,IL-4,IL-6,IL-10,IL-12,tumor necrosis factor(TNF)-α,nuclear factor (NF)-κB and transforming growth factor (TGF)-β Was also enhanced to different degrees.TLR4,a humoral component of innate immune response,increased in all specimens from infected group,whereas Toll-like receptor(TLR)2 expression increased only in one third of these speciraens.The expression of virulent factors including EFG1 (a modulatory factor of hyphal formation),secreted aspartic proteinase (SAP)2,SAP4,SAP5,SAP6,SAP10,lipase(LIP)2,LIP4 and HWP(a major cell wall protein)1 increased in pathogenic strains.Among these differentially expressed genes,monocyte chemoattractant protein(MCP)-1,macrophage inflammatory protein(MIP)-1α,MIP-2,IL-1,TLR4,LIP4 and HWP1,which were involved in extracellular hydrolysis,hyphal formation,phenotypic switching and host native immunity,were increased significantly in all specimens from the infected group.Conclusions Both the deficiency of adaptive immunity and increased virulence of pathogen strains are involved in the pathogenesis of vulvovaginal candidiasis,and TLR4 possibly plays an important role in local immunity of hosts with vulvovaginal candidiasis.