發(fā)布時(shí)間：2022-02-24 07:49

　　目的:本實(shí)驗(yàn)研究高原低氧對(duì)硫酸右旋糖酐鈉（DSS）誘導(dǎo)的潰瘍性結(jié)腸炎小鼠免疫功能的影響。方法:40只Balb/c小鼠隨機(jī)分為四組,分別為:常氧對(duì)照組、DSS誘導(dǎo)產(chǎn)生潰瘍性結(jié)腸炎的常氧DSS組、低氧對(duì)照組和低氧DSS組,常氧組飼養(yǎng)于西寧（海拔2260 m,大氣壓582 mm Hg）;低氧組飼養(yǎng)于模擬海拔5000 m的實(shí)驗(yàn)動(dòng)物低壓艙中,低氧處理7天。通過(guò)流式細(xì)胞術(shù)、ELISA和RT-PCR分別檢測(cè)IL-4、IL-10、1L-17、TNF-α和INF-γ的含量及表達(dá)水平;結(jié)腸組織H.E.染色后觀察其形態(tài)和病理變化。結(jié)果:成功建立了低壓低氧條件下的潰瘍性結(jié)腸炎小鼠模型。實(shí)驗(yàn)結(jié)果表明,小鼠暴露于低壓低氧條件下時(shí),其潰瘍性結(jié)腸炎的表現(xiàn)明顯加重,包括體重減輕、結(jié)腸長(zhǎng)度縮短、同時(shí)伴隨著血便和腹瀉。低氧DSS組的小鼠脾臟和腸系膜淋巴結(jié)中炎癥因子TNF-α、IFN-γ、IL-17和IL-10的濃度顯著高于常氧對(duì)照組,并且結(jié)腸中上述細(xì)胞因子m RNA的表達(dá)量也明顯高于對(duì)照組。此外,結(jié)腸組織髓過(guò)氧化物酶活性顯著增加,組織病理學(xué)分析表明,與常氧對(duì)照組相比低氧增強(qiáng)了DSS誘導(dǎo)的小鼠潰瘍性結(jié)腸炎的嚴(yán)重程度。結(jié)論:高... 

【文章來(lái)源】：青海大學(xué)青海省211工程院校

【文章頁(yè)數(shù)】：68 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
摘要
Abstract
Abbreviations
Chapter 1:Introduction
    1.1 What Is Ulcerative Colitis?
    1.2 Prevalence of ulcerative colitis
    1.3 Etiology of ulcerative colitis
    1.4 Disorder of the immune system
    1.5 DSS model
    1.6 Role of high-altitude hypoxia on immune function
Chapter 2: Materials and Methods
    2.1 Experimental design
    2.2 Mice
    2.3 Chemicals
    2.4 Induction of Ulcerative colitis (UC)
    2.5 Hypoxia induction
    2.6 Clinical markers
        2.6.1 Bodyweight measurement
        2.6.2 Disease activity index (DAI)
        2.6.3 Liquid consumption
        2.6.4 Length of colons
    2.7 Histopathological examinations
        2.7.1 Preparation of colon Cryosections
        2.7.2 Cryosectioning
        2.7.3 Hematoxylin and Eosin(H.E.) Staining
    2.8 Measurement of Myeloperoxidase Activity
    2.9 Enzyme-Linked Immunosorbent Assay (ELISA)
    2.10 Isolation of lymphocytes and purification of CD4+ T cells
    2.11 Flow cytometry assay
    2.12 Gene expression assay
        2.12.1 Quantitative-polymerase chain reaction (Q-PCR) assay
        2.12.2 Isolation of RNA from tissues
        2.12.3 Genome DNA elimination reaction
        2.12.4 Reverse Transcription
        2.12.5 Real-time PCR
    2.13 Statistical analysis
Chapter 3:Results
    3.1 Clinical Features
        3.1.1 Bodyweight measurement
        3.1.2 Liquid consumption
        3.1.3 Disease activity index(DAI)
        3.1.4 Colon Length
    3.2 INFLAMMATORY MARKERS
        3.2.1 Measurement of Myeloperoxidase Activity
        3.2.2 INF-γ
        3.2.3 IL-17
        3.2.4 IL-4
        3.2.5 IL-10
        3.2.6 TNF-α
    3.3 Effect of high-altitude hypoxia on CD4~+ subtypes cells
    3.4 Effect of high-altitude hypoxia on mRNA expression
    3.5 Histological examination
Chapter 4:Discussion
Conclusion and future work
Reference
Acknowledgments
SELF-INTRODUCTION



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