【作者】 张晟瑜；

【导师】 张抒扬； 朱文玲；

【作者基本信息】 北京协和医学院 ， 临床医学， 2010， 博士

【摘要】 研究背景内皮功能紊乱、脂质代谢异常及炎症反应在冠状动脉粥样硬化发生过程中相互作用,形成极其复杂的病理生理学过程。非对称性二甲基精氨酸(ADMA)是内皮型一氧化氮合酶的内源性竞争性抑制剂,它使内皮一氧化氮合成减少,造成内皮功能紊乱。内皮功能紊乱先于冠状动脉粥样硬化发生,并可能导致冠状动脉粥样硬化发生和进展。近年研究发现ADMA与原发性高血压病、高脂血症、糖尿病及高同型半胱氨酸血症等心血管疾病的危险因素相关,但中国人群的ADMA与冠心病预后关系较全面的研究较少。研究目的1、建立高效液相色谱联合串联质谱(HPLC-MS/MS)分析样本ADMA相关指标的方法；2、测定对照组、稳定心绞痛组及急性冠脉综合征组患者血浆样本的ADMA相关指标水平(包括ADMA、对称性二甲基精氨酸(SDMA)、精氨酸(Arg)、瓜氨酸(Cit)、鸟氨酸(Orn)),并探讨其与冠心病及其预后之间的关系。研究方法1、建立HPLC-MS/MS分析样本ADMA相关指标的方法,考察其特异性、敏感度和批内、批间准确度和精密度；并测定入选病例血浆ADMA相关指标水平；2、收集2007年2月至2009年6月,于北京协和医院心内科病房、CCU及急诊住院,行冠状动脉造影术诊断为冠脉正常、稳定心绞痛及急性冠脉综合征患者的血浆样本及临床资料,并随访其2年预后,探讨ADMA相关指标与冠心病及其预后之间的关系。结果1、成功建立HPLC-MS/MS测量ADMA相关指标的方法,并通过方法学考察；2、入选病例共170例,其中对照组20例,稳定心绞痛组45例,急性冠脉综合征组105例(不稳定心绞痛组40例,非ST段抬高型心肌梗死组25例,ST段抬高型心肌梗死组40例),共获得155例随访结果；3、经统计发现ADMA相关指标在冠心病各组间存在明显差异；ADMA水平与TC、 LVEF、BNP等临床及生化指标独立相关；ADMA水平与冠脉病变的范围与严重程度相关；比值Arg/ADMA、Arg+Cit+Orn/ADMA+SDMA是冠心病的独立危险因素；比值Arg/ADMA、Arg+Cit+Orn/ADMA+SDMA是冠心病预后的独立预测因子。结论1、成功建立HPLC-MS/MS分析样本ADMA相关指标的方法；2、ADMA相关指标是冠心病的独立危险因素,且与其预后密切相关。更多还原

【Abstract】 BackgroundEndothelial dysfunction, dyslipidemia and inflammation play critical roles in the process of coronary atherosclerosis. Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of endothelial nitric oxide synthase and an important cause of endothelial dysfunction. Its elevated plasma concentration is associated with a variety of traditional cardiovascular risk factors, and may mediate their effects on vascular endothelium dysfunction. Many studies have implicated ADMA is an independent predictor of coronary artery disease, but rare studies focus on the association between ADMA level and the prognosis of coronary artery disease in Chinese.Objectives1. Establish the high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the analysis of asymmetric dimethylarginine relevant items (including ADMA, SDMA, Arginine, Citrulline and Ornithine) in human plasma.2. Determine the ADMA level of plasma sample from patients in the group of control, stable angina pectoris (SAP) and acute coronary syndrome (ACS) by means of HPLC-MS/MS, and investigate the relationship between ADMA and coronary artery disease and its prognosis.Method1. Build the HPLC-MS/MS method for the analysis of asymmetric dimethylarginine relevant items, and examine its specificity, sensitivity and intra-and inter-batch accuracy and precision.2. Consecutive patients referred to the ward of cardiology department, cardiac care unit and emergency of PUMC for percutaneous coronary intervention from February2007to June2009are enrolled in this study, and their plasma samples and clinical information are collected after informed consent. They are followed up prospectively for2years and the major adverse cardiac events are considered as terminal event. The ADMA levels are determined by HPLC-MS/MS.Results1. Established the HPLC-MS/MS method for the analysis of asymmetric dimethylarginine relevant items successfully and it passed methodology examination.2. A total of170cases are selected, including20cases in control group,45patients in SAP group,105cases in the ACS group, and received follow-up results from155patients.3. The statistics result indicates that significant difference of ADMA relevant items exists among the groups; ADMA concentration correlates with total cholesterol, left ventricle ejection fraction, brain natriuretic peptide and other clinical and biochemical parameters; ADMA concentration is associated with the extent and severity of coronary lesion; ADMA relevant ratios are independent risk factors of coronary disease and independent predict its prognosis.Conclusion1. Establish the HPLC-MS/MS method for the analysis of asymmetric dimethylarginine relevant items.2. ADMA relevant items are independent risk factors of coronary disease, and are closely related to its prognosis.更多还原