高原低氧对DSS诱导小鼠溃疡性结肠炎免疫功能的影响

发布时间：2022-02-24 07:49

　　目的:本实验研究高原低氧对硫酸右旋糖酐钠（DSS）诱导的溃疡性结肠炎小鼠免疫功能的影响。方法:40只Balb/c小鼠随机分为四组,分别为:常氧对照组、DSS诱导产生溃疡性结肠炎的常氧DSS组、低氧对照组和低氧DSS组,常氧组饲养于西宁（海拔2260 m,大气压582 mm Hg）;低氧组饲养于模拟海拔5000 m的实验动物低压舱中,低氧处理7天。通过流式细胞术、ELISA和RT-PCR分别检测IL-4、IL-10、1L-17、TNF-α和INF-γ的含量及表达水平;结肠组织H.E.染色后观察其形态和病理变化。结果:成功建立了低压低氧条件下的溃疡性结肠炎小鼠模型。实验结果表明,小鼠暴露于低压低氧条件下时,其溃疡性结肠炎的表现明显加重,包括体重减轻、结肠长度缩短、同时伴随着血便和腹泻。低氧DSS组的小鼠脾脏和肠系膜淋巴结中炎症因子TNF-α、IFN-γ、IL-17和IL-10的浓度显著高于常氧对照组,并且结肠中上述细胞因子m RNA的表达量也明显高于对照组。此外,结肠组织髓过氧化物酶活性显着增加,组织病理学分析表明,与常氧对照组相比低氧增强了DSS诱导的小鼠溃疡性结肠炎的严重程度。结论:高... 

【文章来源】：青海大学青海省211工程院校

【文章页数】：68 页

【学位级别】：硕士

【文章目录】：
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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