氢分子对感染嗜水气单胞菌斑马鱼的作用研究

发布时间：2018-04-23 03:34

本文选题：氢气 + 嗜水气单孢菌　；参考：《中央民族大学》2017年硕士论文

【摘要】：氢气(Hydrogen,H2)是近年来发现的一种新型治疗性气体,具有抗氧化、抗炎症、抗凋亡及信号通路调节等多重生物学效应,并且在多种动物疾病模型中得到验证。嗜水气单胞菌(Aeromonas hydrophila)属于弧菌科(Vibrionaceae)气单胞菌属(Aeromonas),是水生环境中一类常见致病性病原菌,可以引发鱼类败血症而导致鱼类爆发性死亡。氢气是否对嗜水气单胞菌感染鱼类具有保护作用,其保护机理如何,尚未见报道。本研究以斑马鱼为模式实验动物,建立嗜水气单孢菌感染斑马鱼模型,研究氢气对嗜水气单胞菌感染斑马鱼后的保护效果,并初步探索其保护机理,为氢气用于嗜水气单胞菌感染的防治提供理论和试验依据。主要研究结果如下:1.用富氢水(hydrogen-rich water,HW)处理受嗜水气单孢菌感染的斑马鱼,统计分析细菌感染后斑马鱼的存活率。结果显示1%和4%浓度HW均能起提高到斑马鱼存活率的作用,其中1%HW效果最佳,能将斑马鱼存活率由51.7%提升至72.5%。2.通过细菌学检测HW作用后斑马鱼体内嗜水气单孢菌的生长情况,结果显示HW处理组菌落总数随时间增长速度在各时间段(6h、12h、24h和48h)低于未处理组。3.进一步探究HW在抗嗜水气单孢菌感染斑马鱼中的作用机制,通过荧光定量PCR检测HW处理后斑马鱼脾脏、肾脏和肝脏中免疫调节因子表达量的变化,结果显示在感染后6h、12h及24h,HW能够显著降低三种组织中促炎细胞因子NF-κB、IL-1β及IL-6的表达,并显著上调三种组织中抗炎细胞因子IL-10的表达。4.通过荧光定量PCR检测HW处理后斑马鱼脾脏、肾脏和肝脏中抗氧化相关因子的表达变化,结果显示在感染后6h、12h及24h,HW能够显著提高三种组织中抗氧化因子SOD1、CAT及POD的表达。本研究发现在嗜水气单孢菌感染斑马鱼时,HW能够降低嗜水气单孢菌的增殖,降低斑马鱼组织促炎细胞因子表达,提高其抗炎细胞因子表达,还能够提高组织抗氧化酶基因的表达,揭示HW可能是通过降低病原菌增殖、调节炎症相关因子的表达,提高抗氧化相关因子的表达,来缓解鱼体炎症反应、抵御嗜水气单孢菌感染,从而为进一步探究氢气的治疗作用提供了坚实基础。
[Abstract]:Hydrogenin H _ 2 is a new therapeutic gas found in recent years. It has many biological effects, such as anti-oxidation, anti-inflammation, anti-apoptosis and signal pathway regulation, and has been verified in many animal disease models. Aeromonas hydrophila belongs to Vibrionaceae.Aeromonas hydrophila belongs to Aeromonas. Aeromonas hydrophila is a common pathogenic bacteria in aquatic environment, which can cause septicemia of fish and lead to sudden death of fish. Whether hydrogen has protective effect on Aeromonas hydrophila infected fish and its protective mechanism has not been reported. In this study, zebrafish were used as model animals to establish a zebrafish model infected by Aeromonas hydrophila, to study the protective effect of hydrogen on zebrafish infected by Aeromonas hydrophila, and to explore its protective mechanism. To provide theoretical and experimental basis for the prevention and treatment of Aeromonas hydrophila infection by hydrogen. The main results are as follows: 1. Zebrafish infected by Aeromonas hydrophila were treated with hydrogen-rich water monosporium (HW) and the survival rate of zebrafish after bacterial infection was analyzed statistically. The results showed that both 1% and 4% HW could increase the survival rate of zebrafish, and 1%HW had the best effect, which could increase the survival rate of zebrafish from 51.7% to 72.5%. The growth of Aeromonas hydrophila in zebrafish treated with HW was detected by bacteriology. The results showed that the total colony number in HW treated group was significantly lower than that in untreated group for 24 h and 48 h after HW treatment. To further explore the mechanism of HW in resisting the infection of zebrafish with Aeromonas hydrophila, the expression of immunomodulators in spleen, kidney and liver of zebrafish treated with HW was detected by fluorescence quantitative PCR. The results showed that HW significantly decreased the expression of NF- 魏 B, IL-1 尾 and IL-6, and upregulated the expression of anti-inflammatory cytokine IL-10. The expression of antioxidant related factors in spleen, kidney and liver of zebrafish treated with HW was detected by fluorescence quantitative PCR. The results showed that HW could significantly increase the expression of SOD1cat and POD in the three tissues at 6h, 12h and 24h after HW infection. It was found that HW could reduce the proliferation of Aeromonas hydrophila, reduce the expression of pro-inflammatory cytokines and increase the expression of anti-inflammatory cytokines in zebrafish tissues when Aeromonas hydrophila infected zebrafish. HW may alleviate the inflammatory response of fish by reducing the proliferation of pathogenic bacteria, regulating the expression of inflammatory factors and increasing the expression of antioxidant related factors. To resist the infection of Aeromonas hydrophila, it provides a solid foundation for further exploring the therapeutic effect of hydrogen.
【学位授予单位】：中央民族大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S941.4

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