强脉冲光420nm对红色毛癣菌的干预作用

发布时间：2018-04-08 14:56

本文选题：红色毛癣菌　切入点：强脉冲光　出处：《南方医科大学》2017年硕士论文

【摘要】：[背景]研究发现光疗对真菌即可产生直接杀灭作用,又可通过诱导其产生大量胞内活性氧簇(Reactive oxygen species,ROS)造成氧化损伤。推测强脉冲光(intense pulsed light,IPL)具有类似作用。在真核生物中还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶(nicotinamideadenine dinucleotide phosphate oxidase,NADPH oxidase,Nox)是诱导细胞活性氧簇的产生的快速反应酶。真菌中NADPH氧化酶上调时,同样可促使ROS产生增加,导致氧化应激损伤。研究发现LED可抑制T.rubrum,引起氧化应激及氮化应激指标显著性上升。因此,我们拟探讨IPL是否通过氮化应激抑制红色毛癣菌(Trichophyton rubrum,T.rubrum)。本研究运用IPL420nm体外干预红色毛癣菌,初步探讨对其影响。[研究目的]通过检测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)表达及角质蛋白酶活力,证实IPL420nm对T.rubrum的抑制作用和探讨其影响,为IPL420nm应用于治疗T.rubrum等浅表真菌感染性疾病提供理论基础。[方法](1)培养红色毛癣菌(临床株和标准株ATCC4438),分2组:T.rubrum,T.rubrum+IPL,分别做能量依赖及时间依赖测试,选取最适参数。(2)实验分3组:T.rubrum,T.rubrum+IPL,T.rubrum+DPI(NADPH 氧化酶抑制剂)+IPL,检测真菌活力、ROS、MDA和SOD、GSH-PX、角蛋白酶水平及NADPH氧化酶表达,扫描电镜观测真菌表面变化。(3)实验分3组:T.rubrum,T.rubrum+IPL,T.rubrum+L-NMMA(总一氧化氮合成酶抑制剂)+IPL,检测真菌活力、NO、NOS及角蛋白酶水平,扫描电镜观测真菌表面变化。[结果](1)IPL对临床株及ATCC4438标准株的生长抑制作用呈能量剂量依赖和时间依赖。(2)IPL处理后,真菌结构损伤严重,ROS及MDA含量增加,SOD、GSH-PX和角蛋白酶活性下降,Nox基因表达上调,DPI可以抑制IPL的作用。(3)IPL处理后,真菌结构损伤严重,NO水平增加,角蛋白酶活性下降,NOS表达上调,而L-NMMA可以抑制IPL的作用。[结论](1)通过IPL420nm诱导产生ROS和NO,导致氧化和氮化损伤,抑制T.rubrum生长,降低真菌毒力;(2)IPL420nm可能通过NADPH氧化酶和一氧化氮合成酶抑制T.rubrum;(3)为强脉冲光可能成为一种治疗浅表真菌感染性疾病安全的新方法提供理论依据。
[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.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R756

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