本文選題：紅色毛癬菌　切入點(diǎn)：強(qiáng)脈沖光　出處：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[背景]研究發(fā)現(xiàn)光療對真菌即可產(chǎn)生直接殺滅作用,又可通過誘導(dǎo)其產(chǎn)生大量胞內(nèi)活性氧簇(Reactive oxygen species,ROS)造成氧化損傷。推測強(qiáng)脈沖光(intense pulsed light,IPL)具有類似作用。在真核生物中還原型煙酰胺腺嘌呤二核苷酸磷酸氧化酶(nicotinamideadenine dinucleotide phosphate oxidase,NADPH oxidase,Nox)是誘導(dǎo)細(xì)胞活性氧簇的產(chǎn)生的快速反應(yīng)酶。真菌中NADPH氧化酶上調(diào)時(shí),同樣可促使ROS產(chǎn)生增加,導(dǎo)致氧化應(yīng)激損傷。研究發(fā)現(xiàn)LED可抑制T.rubrum,引起氧化應(yīng)激及氮化應(yīng)激指標(biāo)顯著性上升。因此,我們擬探討IPL是否通過氮化應(yīng)激抑制紅色毛癬菌(Trichophyton rubrum,T.rubrum)。本研究運(yùn)用IPL420nm體外干預(yù)紅色毛癬菌,初步探討對其影響。[研究目的]通過檢測IPL抑制T.rubrum中的ROS和NO水平、NADPH氧化酶和一氧化氮合成酶(NOS)表達(dá)及角質(zhì)蛋白酶活力,證實(shí)IPL420nm對T.rubrum的抑制作用和探討其影響,為IPL420nm應(yīng)用于治療T.rubrum等淺表真菌感染性疾病提供理論基礎(chǔ)。[方法](1)培養(yǎng)紅色毛癬菌(臨床株和標(biāo)準(zhǔn)株ATCC4438),分2組:T.rubrum,T.rubrum+IPL,分別做能量依賴及時(shí)間依賴測試,選取最適參數(shù)。(2)實(shí)驗(yàn)分3組:T.rubrum,T.rubrum+IPL,T.rubrum+DPI(NADPH 氧化酶抑制劑)+IPL,檢測真菌活力、ROS、MDA和SOD、GSH-PX、角蛋白酶水平及NADPH氧化酶表達(dá),掃描電鏡觀測真菌表面變化。(3)實(shí)驗(yàn)分3組:T.rubrum,T.rubrum+IPL,T.rubrum+L-NMMA(總一氧化氮合成酶抑制劑)+IPL,檢測真菌活力、NO、NOS及角蛋白酶水平,掃描電鏡觀測真菌表面變化。[結(jié)果](1)IPL對臨床株及ATCC4438標(biāo)準(zhǔn)株的生長抑制作用呈能量劑量依賴和時(shí)間依賴。(2)IPL處理后,真菌結(jié)構(gòu)損傷嚴(yán)重,ROS及MDA含量增加,SOD、GSH-PX和角蛋白酶活性下降,Nox基因表達(dá)上調(diào),DPI可以抑制IPL的作用。(3)IPL處理后,真菌結(jié)構(gòu)損傷嚴(yán)重,NO水平增加,角蛋白酶活性下降,NOS表達(dá)上調(diào),而L-NMMA可以抑制IPL的作用。[結(jié)論](1)通過IPL420nm誘導(dǎo)產(chǎn)生ROS和NO,導(dǎo)致氧化和氮化損傷,抑制T.rubrum生長,降低真菌毒力;(2)IPL420nm可能通過NADPH氧化酶和一氧化氮合成酶抑制T.rubrum;(3)為強(qiáng)脈沖光可能成為一種治療淺表真菌感染性疾病安全的新方法提供理論依據(jù)。
[Abstract]:[background] it has been found that phototherapy can not only kill fungi directly, but also cause oxidative damage by inducing them to produce a large number of reactive oxygen speciesros.It is speculated that intense pulsed lightlight (IPL) has a similar effect.In eukaryotes, nicotinamide adenine dinucleotide phosphate oxidase (nicotinamideadenine dinucleotide phosphate oxidase Nox) is a rapid reactive enzyme that induces the production of reactive oxygen species.When NADPH oxidase was up-regulated in fungi, ROS production was also increased, resulting in oxidative stress injury.It was found that LED could inhibit T. rubrum and induce a significant increase in oxidative stress and nitrided stress.Therefore, we intend to investigate whether IPL inhibits Trichophyton rubrumus T. rubrumus by nitridation stress.In this study, IPL420nm was used to interfere with Trichophyton rubrum in vitro.[objective] to investigate the inhibitory effect of IPL420nm on T.rubrum by detecting the inhibitory effect of IPL on the expression of ROS and nitric oxide oxidase (NOS) and nitric oxide synthase (NOS) and the activity of keratinase in T.rubrum.To provide a theoretical basis for the application of IPL420nm in the treatment of superficial fungal infections such as T.rubrum.[methods] cultured Trichophyton rubrum (clinical strain and standard strain ATCC 4438) were divided into two groups: T. rubrumum T. rubrum IPL, and were tested for energy dependence and time dependence, respectively.Select the optimum parameter. The experiment was divided into three groups: T. T. rubrum.T. rubrum DPI(NADPH oxidase inhibitor). The activity of fungi was detected by Ros MDA, SOD GSH-PX, keratinase level and NADPH oxidase expression.The surface changes of fungi were observed by scanning electron microscope (SEM). The experiment was divided into 3 groups: T. T. rubrumum T. rubrum IPL T. rubrum L-NMMA-IPL (total nitric oxide synthase inhibitor) IPL. the NON-NOS and keratinase levels of fungi were detected, and the surface changes of fungi were observed by scanning electron microscope (SEM).[results] the inhibitory effects of ATCC4438 on the growth of clinical and ATCC4438 standard strains were treated in a dose-and time-dependent manner.The contents of Ros and MDA increased in severe structural damage of fungi. The decreased activity of GSH-PX and keratinase. The up-regulated expression of Nox gene could inhibit the effect of IPL. The level of no was increased and the activity of keratinase was decreased.L-NMMA can inhibit the effect of IPL.[conclusion] IPL420nm induces the production of ROS and no, resulting in oxidative and nitrided damage and inhibiting the growth of T.rubrum.The reduction of fungal virulence, IPL 420 nm, which may inhibit T. rubrumin3 by NADPH oxidase and nitric oxide synthase, may be a new safe method for the treatment of superficial fungal infectious diseases.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R756

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