儿茶酚胺类激素对外周细胞炎性因子表达影响的研究

论文摘要

第一部分 研究背景：动脉粥样硬化（artherosclerosis，As）病程复杂，涉及到内皮损伤，粘附分子表达增强，血小板附着血管壁，单核淋巴细胞粘着渗入到动脉壁，促炎细胞因子出现，LDL的氧化和浸润，单核细胞增生、迁移、活化，巨噬细胞和平滑肌泡沫化等。目前对As发病机理已提出众多学说，主要有脂肪浸润学说、血小板聚集和血栓形成学说、平滑肌细胞克隆学说和损伤反应学说。近年来研究发现，As的病理表现具有炎症病理的基本表现形式，随着炎症细胞和炎症介质在动脉粥样硬化斑块中的不断检出，越来越多的研究结果支持As不单纯是内皮损伤后被动的脂质浸润或修复过程，更重要的是一种活跃的炎症性疾病。许多证据表明心理应激是心血管疾病的危险因子，心理社会因素参与一部分心血管疾病如As的起病和进程，心血管疾病与心理社会因素之间的关联日益引起重视，但其生物学机制并不十分清楚。 应激是内环境稳态受到威胁的状态，当机体受到环境、心理、生理性应激原等的刺激时均可以引起机体的应激反应，激活交感神经系统（sympathetic nervous system，SNS）和下丘脑-垂体-肾上腺轴（hypothalamic-pituitary-adrenal axis，HPA）产生相应的生理学变化。在易感者或高危人群中，反复的急性应激或者慢性应激激活SNS、HPA等可引起儿茶酚胺类激素、皮质类固醇等一系列的应激性激素的释放，结果导致心血管反应性增高，改变血流和血压导致内皮损伤和血小板附壁；细胞因子和其它因子诱导功能异常的内皮细胞表达粘附分子，募集单核淋巴细胞粘着进入内膜。反复的急性应激或者慢性应激也可激活单核细胞、肥大细胞等，产生细胞因子和炎性介质等可加重炎症反应。即应激时的皮质类固醇和儿茶酚胺类激素实际上启动了和炎症反应相似的调节机制，如生成细胞因子和急性期反应物质，心理性刺激触发的应激反应涵盖了炎性反

论文目录

• Abstract in Chinese
• Abstract in English
• Part One: Research on Catecholamine Hormone's Effect on Expression of the Peripheral Cellular Inflammatory Factor
• Introduction
• 1 Materials
• 1 1 Main reagents
• 1.2 Instrumenttations
• 2 Empirical Method
• 2.1 Cell culture
• 2.2 Extraction of total RNA and purification of mRNA
• 2.3 Quantitation and integrality analysis on RNA sample
• 2.4 Suppression subtractive hybridization （SSH）
• 2.5 PCR analysis of subtraction efficiency
• 2.6 Construct differential expression gene of subtractive cDNA Lib
• 2.7 Screening subtractive cDNA Lib by reverse dot blot hybridization
• 2.8 DNA sequencing and analysis of homology
• 2.9 Rverse transcription-polymerase chain reaction
• 2.10 Detection of IL-6 protein expression by ELISA
• 2.11 Statistical analyses
• 3 Results
• 3.1 Abstraction of total RNA and mRNA
• 3.2 Synthesis of ds cDNA and the result of Rsal digestion
• 3.3 Results of the ligation efficiency analysis
• 3.4 Two series of PCR on differential hybridization product
• 3.5 Results of subtractive hybridization efficiency analysis
• 3.6 Construction of subtractive cDNA Lib
• 3.7 Propagation of positive colonies
• 3.8 Bolting of subtractive cDNA Lib by PCR
• 3.9 The result of reverse dot blot hybridization
• 3.10 DNA sequencing and homology analytic results of positive colonies
• 3.11 The effect of adrenalin on the expression of inflammatory factors of cultured cells
• 3.12 The effect of noradrenalin exposed to cultured cells on the expression of inflammatory factors
• 3.13 The effect of cortisol exposed to cultured cells on the expression of inflammatory factors
• 4 Discussion
• 5 Conclusion
• 6 References
• Part Two: Relationship of APOE Gene Polymorphism with Subclasses of Plasma High Density Lipoprotein in Hyperlipidemia
• Introduction
• 1 Subjects and methods
• 1.1 Subjects
• 1.2 Specimens
• 1.3 Genetic analyses
• 1.4 Plasma lipid and apolipoprotein analyses
• 1.5 HDL subclasses analyses
• 1.6 Statistical analyses
• 2 Results
• 2.1 Frequency distributions of APOE genotypes and alleles in hyperlipidemic and control groups
• 2.2 Concentrations of plasma lipids, lipoproteins, apolipoproteins and relative APOAI percentage contents of plasma HDL subclasses
• 2.3 Lipid parameters and relative APOAI percentage contents of plasma HDL subclasses with different APOE genotypes
• 3 Discussion
• 4 Conclusion
• 5 References
• Part Three: Relationship between Plasma HDL Subclasses Distribution and Lipoprotein Lipase Gene HindⅢ Polymorphism in Hyperlipidemia
• Introduction
• 1 Subjects and methods
• 1.1 Subjects
• 1.2 Specimens
• 1.3 Plasma lipid and apolipoprotein analyses
• 1.4 HDL subclasses analyses
• 1.5 Genetic analyses
• 1.6 Statistical analyses
• 2 Results
• 2.1 Frequency distributions of LPL genotypes and alleles in the hyperlipidemic and control group
• 2.2 Demographic characteristics, lipid parameters and relative apoA-I contents of plasma HDL subclasses
• 2.3 Lipid parameters and relative apoA-I contents of plasma HDL subclasses with different LPL genotypes
• 3 Discussion
• 4 Conclusion
• 5 References
• Review
• Brief introduction of technological research findings during study
• The statement of thesis' originality
• Acknowledgements