犬小孢子菌FSH1在儿童头皮与光滑皮肤组织诱导前后的表达

发布时间：2018-05-16 20:16

本文选题：犬小孢子菌 + FSH1　；参考：《大连医科大学》2010年硕士论文

【摘要】： 目的：犬小孢子菌是亲动物性真菌,常常感染动物(尤其是猫狗)及人类,导致真菌性疾病。它易于感染头皮组织,且以儿童多见。近年来,人类与猫狗等宠物的密切接触导致犬小孢子菌病发病率逐年上升。在研究犬小孢子菌致病作用过程中,蛋白酶类与头癣的相关性及在头癣的发病机制中的作用是研究关注的重点。本实验应用半定量RT—PCR技术,检测30株犬小孢子菌头癣株和30株体癣株的丝氨酸水解酶(FSH1)表达水平；并检测二者于沙堡液体培养基传代后FSH1的表达及分别经儿童头皮、儿童光滑皮肤诱导前后FSH1的表达。期望本实验结果能为FSH1功能分析奠定一定基础,为FSH1在犬小孢子菌所致头癣发病机制中的作用提供一定的依据。方法：30株头癣株和30株体癣株接种于米饭培养基中,培养2周,分离孢子。将5×106个孢子接种于100ml沙堡液体培养基,培养21-28天后收集培养基中的菌丝团,为第一代,用同样的方法传五代后得第五代。同时将头癣株和体癣株各5×106个孢子分别接种于100ml儿童头皮诱导培养基和100ml儿童光滑皮肤诱导培养基,培养21-28天后收集两种皮肤组织诱导培养基中的菌丝团。应用半定量RT—PCR检测以上各菌株的FSH1mRNA表达水平。结果：犬小孢子菌在沙堡液体培养基中及含儿童头皮组织和儿童光滑皮肤两种底物诱导培养基中均生长,形成菌丝团块。该菌在沙堡液体培养基中生长较佳,在含儿童头皮组织和儿童光滑皮肤组织的诱导培养基中生长略差。应用半定量RT—PCR检测犬小孢子菌的FSH1mRNA表达水平。结果发现FSH1在犬小孢子菌中都有表达,且头癣株和体癣株FSH1mRNA表达水平差异明显(P0.01)；两者于沙堡液体培养基传代后FSH1mRNA表达水平无明显差异(P0.05)；皮肤组织诱导前后的菌株FSH1mRNA表达水平差异明显(P0.01),且儿童头皮、儿童光滑皮肤诱导后菌株FSH1mRNA表达水平差异明显(P0.01)。结论：1.FSH1在犬小孢子菌中有活性表达。2.头癣株FSH1mRNA表达量明显高于体癣株。3.经沙堡液体培养基传代的菌株能稳定表达FSH1,且不受传代影响。4.经皮肤组织诱导后,FSH1mRNA表达水平明显升高,且儿童头皮组织诱导后的菌株其表达最明显。5.FSH1可能与犬小孢子菌致头癣有关。
[Abstract]:Objective: microsporozoa canis is a zoophile fungus, which often infects animals (especially cats and dogs) and humans, leading to fungal diseases. It is easy to infect scalp tissue, and is more common in children. In recent years, the close contact between human beings and pets, such as cats and dogs, has led to an increase in the incidence of microsporosis in dogs. The relationship between proteinases and tinea capitis and its role in the pathogenesis of tinea capitis is the focus in the study of pathogenicity of microsporum canis. The expression of serine hydrolase (FSH1) in 30 strains of Trichophyton canis and 30 strains of tinea corporis was detected by semi-quantitative RT-PCR technique, and the expression of FSH1 was detected after the passage of the two strains in the liquid medium of Shabourg and the scalp of children, respectively, and the expression of FSH1 in 30 strains of Tinea canis and 30 strains of tinea corporis was detected. Expression of FSH1 in children with smooth skin before and after induction. It is expected that the results of this study can lay a foundation for the functional analysis of FSH1 and provide a basis for the role of FSH1 in the pathogenesis of tinea capitis induced by microsporozoa canis. Methods 30 strains of tinea capitis and 30 strains of tinea corporis were inoculated in rice medium and cultured for 2 weeks. 5 脳 106 spores were inoculated on 100ml sand castle liquid medium. The hyphae were collected from the culture medium 21-28 days later. The first generation was obtained by the same method. The fifth generation was obtained after the fifth generation was passed on by the same method. At the same time, 5 脳 106 spores of tinea capitis and tinea corporis were inoculated into 100ml children's scalp induction medium and 100ml child smooth skin induction medium respectively. The expression level of FSH1mRNA was detected by semi-quantitative RT-PCR. Results: Microsporum canis grew in the liquid medium of Shabourg and in the substrates containing children's scalp tissue and smooth skin of children, forming hyphal mass. The strain grew better in the liquid medium of Shabourg, and grew slightly worse in the induction medium containing the scalp tissue of children and the smooth skin tissue of the children. The expression of FSH1mRNA in microsporozoa canis was detected by semi-quantitative RT-PCR. The results showed that FSH1 was expressed in microsporum canis. The difference of FSH1mRNA expression between tinea capitis and tinea corporis was significant (P 0.01), there was no significant difference in FSH1mRNA expression between the two groups after the passage of Shabao liquid medium, the expression level of FSH1mRNA was significantly different between the two strains before and after induction of skin tissue, and the scalp of children. The expression level of FSH1mRNA in children with smooth skin was significantly different (P 0.01). Conclusion: 1. FSH1 has active expression in microsporum canis. The expression of FSH1mRNA in tinea capitis was significantly higher than that in tinea corporis. The strain subcultured in Shabourg liquid medium could express FSH1 stably and was not affected by passage. 4. The expression level of FSH1 mRNA was significantly increased after skin induction, and the expression of FSH1 was the most obvious in children's scalp tissue induced. 5. FSH1 may be related to the tinea capitis induced by microsporozoa canis.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R756

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