本文選題：犬小孢子菌 + FSH1　； 參考：《大連醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的：犬小孢子菌是親動(dòng)物性真菌,常常感染動(dòng)物(尤其是貓狗)及人類(lèi),導(dǎo)致真菌性疾病。它易于感染頭皮組織,且以兒童多見(jiàn)。近年來(lái),人類(lèi)與貓狗等寵物的密切接觸導(dǎo)致犬小孢子菌病發(fā)病率逐年上升。在研究犬小孢子菌致病作用過(guò)程中,蛋白酶類(lèi)與頭癬的相關(guān)性及在頭癬的發(fā)病機(jī)制中的作用是研究關(guān)注的重點(diǎn)。本實(shí)驗(yàn)應(yīng)用半定量RT—PCR技術(shù),檢測(cè)30株犬小孢子菌頭癬株和30株體癬株的絲氨酸水解酶(FSH1)表達(dá)水平；并檢測(cè)二者于沙堡液體培養(yǎng)基傳代后FSH1的表達(dá)及分別經(jīng)兒童頭皮、兒童光滑皮膚誘導(dǎo)前后FSH1的表達(dá)。期望本實(shí)驗(yàn)結(jié)果能為FSH1功能分析奠定一定基礎(chǔ),為FSH1在犬小孢子菌所致頭癬發(fā)病機(jī)制中的作用提供一定的依據(jù)。 方法：30株頭癬株和30株體癬株接種于米飯培養(yǎng)基中,培養(yǎng)2周,分離孢子。將5×106個(gè)孢子接種于100ml沙堡液體培養(yǎng)基,培養(yǎng)21-28天后收集培養(yǎng)基中的菌絲團(tuán),為第一代,用同樣的方法傳五代后得第五代。同時(shí)將頭癬株和體癬株各5×106個(gè)孢子分別接種于100ml兒童頭皮誘導(dǎo)培養(yǎng)基和100ml兒童光滑皮膚誘導(dǎo)培養(yǎng)基,培養(yǎng)21-28天后收集兩種皮膚組織誘導(dǎo)培養(yǎng)基中的菌絲團(tuán)。應(yīng)用半定量RT—PCR檢測(cè)以上各菌株的FSH1mRNA表達(dá)水平。 結(jié)果：犬小孢子菌在沙堡液體培養(yǎng)基中及含兒童頭皮組織和兒童光滑皮膚兩種底物誘導(dǎo)培養(yǎng)基中均生長(zhǎng),形成菌絲團(tuán)塊。該菌在沙堡液體培養(yǎng)基中生長(zhǎng)較佳,在含兒童頭皮組織和兒童光滑皮膚組織的誘導(dǎo)培養(yǎng)基中生長(zhǎng)略差。應(yīng)用半定量RT—PCR檢測(cè)犬小孢子菌的FSH1mRNA表達(dá)水平。結(jié)果發(fā)現(xiàn)FSH1在犬小孢子菌中都有表達(dá),且頭癬株和體癬株FSH1mRNA表達(dá)水平差異明顯(P0.01)；兩者于沙堡液體培養(yǎng)基傳代后FSH1mRNA表達(dá)水平無(wú)明顯差異(P0.05)；皮膚組織誘導(dǎo)前后的菌株FSH1mRNA表達(dá)水平差異明顯(P0.01),且兒童頭皮、兒童光滑皮膚誘導(dǎo)后菌株FSH1mRNA表達(dá)水平差異明顯(P0.01)。 結(jié)論：1.FSH1在犬小孢子菌中有活性表達(dá)。2.頭癬株FSH1mRNA表達(dá)量明顯高于體癬株。3.經(jīng)沙堡液體培養(yǎng)基傳代的菌株能穩(wěn)定表達(dá)FSH1,且不受傳代影響。4.經(jīng)皮膚組織誘導(dǎo)后,FSH1mRNA表達(dá)水平明顯升高,且兒童頭皮組織誘導(dǎo)后的菌株其表達(dá)最明顯。5.FSH1可能與犬小孢子菌致頭癬有關(guān)。
[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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R756

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