中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (4): 238-242.

• 论著 • 上一篇    下一篇

须癣毛癣菌肉芽肿株与体癣株形态学、致病性及比较蛋白质组学的差异性研究

王静1,杨国玲2   

  1. 1. 江苏省徐州市儿童医院皮肤科
    2. 大连医科大学附属第一医院
  • 收稿日期:2011-05-23 修回日期:2011-12-14 出版日期:2012-04-15 发布日期:2012-03-30
  • 通讯作者: 杨国玲 E-mail:Yanggl@medmail.com.cn

Comparison of morphology, pathogenicity and proteomics of granuloma- and tinea corporis-derived Trichophyton mentagrophytes

Jing WANG,yang guoling   

  • Received:2011-05-23 Revised:2011-12-14 Online:2012-04-15 Published:2012-03-30
  • Contact: yang guoling E-mail:Yanggl@medmail.com.cn

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摘要：

目的 筛选须癣毛癣菌肉芽肿株和体癣株的差异表达蛋白，探讨须癣毛癣菌致深部和浅部感染的发病机制。方法 选取4株肉芽肿株和4株体癣株须癣毛癣菌，行27 ℃和37 ℃平皿及钢圈法小培养，观察形态变化。取8只豚鼠，糖皮质激素干预改变豚鼠的免疫状态，每只豚鼠对称部位分别采用表面和皮下两种接种方式接种同一种菌，制备体癣模型和肉芽肿模型，接种后2周左右直接镜检及培养、组织病理检查，验证感染效果。筛选肉芽肿株强毒株和体癣株弱毒株各1株，行双向电泳，质谱检测和NCBI数据库分析。结果 须癣毛癣菌37 ℃培养，4株肉芽肿株较4株体癣株生长良好；27 ℃培养，两者形态差别不明显。豚鼠感染实验，所有菌株均构建了体癣模型，4株肉芽肿株较4株体癣株炎症反应明显。3株肉芽肿株形成皮下结节，直接镜检及培养、组织病理检查均显示感染成功。比较蛋白质组学，肉芽肿株和体癣株分别检测到蛋白点（463 ± 20）个与（398 ± 17）个，肉芽肿株有62个蛋白上调表达，体癣株有21个蛋白上调表达，肉芽肿株中有致病意义的蛋白包括烯醇化酶、热休克蛋白家族、丝-苏氨酸蛋白酶、细胞信号转导相关蛋白、能量相关蛋白、细胞骨架蛋白及一些假想蛋白。结论 须癣毛癣菌肉芽肿株较体癣株耐热，易感染宿主；两者蛋白质水平有明显差异。

关键词: 毒力

Abstract:

Objective To screen differentially expressed proteins between granuloma- and tinea corporis-derived Trichophyton mentagrophytes strains, and to explore the pathogenesis of T. mentagrophytes in deep and superficial infection. Methods Four T. mentagrophytes isolates from granuloma and 4 isolates from tinea corporis were cultured in agar plates and small steel rings at 27 ℃ and 37 ℃ respectively, followed by morphologic observation. Eight guinea pigs were immunocompromised by glucocorticoids, and superficially and subcutaneously inoculated with the same amount of fungal suspension to develop an animal model of tinea corporis and granuloma, respectively. Two weeks later, the infection of guinea pigs was confirmed by microscopy, fungal culture and histopathology. Proteins were extracted from a highly toxic granuloma-derived T. mentagrophytes strain and a lowly toxic tinea corporis-derived T. mentagrophytes strain, and subjected to two-dimensional electrophoresis, mass spectrometry and identification by using the National Center for Biotechnology Information （NCBI） database. Results The granuloma-derived T. mentagrophytes isolates grew better at 37 ℃ than tinea corporis-derived T. mentagrophytes isolates did, while no significant difference was observed in the morphology of colonies between the two kinds of T. mentagrophytes isolates at 27 ℃. Tinea corporis models were successfully established in guinea pigs with the 8 T. mentagrophytes strains, and the granuloma-derived isolates induced a more intense inflammation than tinea corporis-derived isolates. Granuloma model was constructed only with 3 granuloma-derived strains, which was proved by microscopy, fungal culture and histopathology. A total of 463 ± 20 and 398 ± 17 protein spots were detected, with 62 and 21 upregulated proteins, from granuloma-derived and tinea corporis-derived T. mentagrophytes strains, respectively. Bioinformatics analysis revealed the following meaningful proteins from differentially expressed proteins in granuloma-derived T. mentagrophytes strains, including enolase, heat shock protein, serine/threonine protease, cellular signal transduction proteins, energy metabolism-related proteins, cytoskeleton proteins and some hypothetical proteins. Conclusions The granuloma-derived T. mentagrophytes strains are more heat-resistant with a higher potency to cause infection compared with tinea corporis-derived T. mentagrophytes strains. Differences exist in the expression of proteins between granuloma- and tinea corporis-derived T. mentagrophytes strains.