本文選題：氫氣 + 嗜水氣單孢菌　； 參考：《中央民族大學(xué)》2017年碩士論文

【摘要】：氫氣(Hydrogen,H2)是近年來發(fā)現(xiàn)的一種新型治療性氣體,具有抗氧化、抗炎癥、抗凋亡及信號(hào)通路調(diào)節(jié)等多重生物學(xué)效應(yīng),并且在多種動(dòng)物疾病模型中得到驗(yàn)證。嗜水氣單胞菌(Aeromonas hydrophila)屬于弧菌科(Vibrionaceae)氣單胞菌屬(Aeromonas),是水生環(huán)境中一類常見致病性病原菌,可以引發(fā)魚類敗血癥而導(dǎo)致魚類爆發(fā)性死亡。氫氣是否對(duì)嗜水氣單胞菌感染魚類具有保護(hù)作用,其保護(hù)機(jī)理如何,尚未見報(bào)道。本研究以斑馬魚為模式實(shí)驗(yàn)動(dòng)物,建立嗜水氣單孢菌感染斑馬魚模型,研究氫氣對(duì)嗜水氣單胞菌感染斑馬魚后的保護(hù)效果,并初步探索其保護(hù)機(jī)理,為氫氣用于嗜水氣單胞菌感染的防治提供理論和試驗(yàn)依據(jù)。主要研究結(jié)果如下:1.用富氫水(hydrogen-rich water,HW)處理受嗜水氣單孢菌感染的斑馬魚,統(tǒng)計(jì)分析細(xì)菌感染后斑馬魚的存活率。結(jié)果顯示1%和4%濃度HW均能起提高到斑馬魚存活率的作用,其中1%HW效果最佳,能將斑馬魚存活率由51.7%提升至72.5%。2.通過細(xì)菌學(xué)檢測(cè)HW作用后斑馬魚體內(nèi)嗜水氣單孢菌的生長(zhǎng)情況,結(jié)果顯示HW處理組菌落總數(shù)隨時(shí)間增長(zhǎng)速度在各時(shí)間段(6h、12h、24h和48h)低于未處理組。3.進(jìn)一步探究HW在抗嗜水氣單孢菌感染斑馬魚中的作用機(jī)制,通過熒光定量PCR檢測(cè)HW處理后斑馬魚脾臟、腎臟和肝臟中免疫調(diào)節(jié)因子表達(dá)量的變化,結(jié)果顯示在感染后6h、12h及24h,HW能夠顯著降低三種組織中促炎細(xì)胞因子NF-κB、IL-1β及IL-6的表達(dá),并顯著上調(diào)三種組織中抗炎細(xì)胞因子IL-10的表達(dá)。4.通過熒光定量PCR檢測(cè)HW處理后斑馬魚脾臟、腎臟和肝臟中抗氧化相關(guān)因子的表達(dá)變化,結(jié)果顯示在感染后6h、12h及24h,HW能夠顯著提高三種組織中抗氧化因子SOD1、CAT及POD的表達(dá)。本研究發(fā)現(xiàn)在嗜水氣單孢菌感染斑馬魚時(shí),HW能夠降低嗜水氣單孢菌的增殖,降低斑馬魚組織促炎細(xì)胞因子表達(dá),提高其抗炎細(xì)胞因子表達(dá),還能夠提高組織抗氧化酶基因的表達(dá),揭示HW可能是通過降低病原菌增殖、調(diào)節(jié)炎癥相關(guān)因子的表達(dá),提高抗氧化相關(guān)因子的表達(dá),來緩解魚體炎癥反應(yīng)、抵御嗜水氣單孢菌感染,從而為進(jìn)一步探究氫氣的治療作用提供了堅(jiān)實(shí)基礎(chǔ)。
[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.
【學(xué)位授予單位】：中央民族大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S941.4

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