冠心病心外膜脂肪微环境变化对局部脂联素糖基化修饰的影响及其机制研究

The earlier research of this project discovered that it has formed atherosclerotic plaques in coronary artery, but not easy to occur in internal mammary artery and radial artery under identical macroenvironment in coronary heart disease, moreover, perivascular adipose tissue of lesion of coronary artery was endocrine dysfunction and endoplasmic reticulum stress. This shows that local micro-environment of the heart affects coronary artery disease(CAD). In coronary disease animal model, apM1 gene was local transferred to epicardial adipose to Anti-atherosclerosis, results is ineffective. Follow-up study found that under the condition of endoplasmic reticulum stress, post-translational process of adiponectin was blocked. Increase in mRNA levels didn't lead to a corresponding increase in the level of adiponectin. According to the above, from the perspective of post-translational modification, this project further study the rule of glycosylation of adiponectin in local micro-environment of CAD at In Vivo, subcell and animal model level. Using the methods of SDS/PAGE and mass spectrum precise analyze the changes of glycosylation site of adiponectin, and study the effect and mechanism of adiponectin protein of various post-translational stages in apoE-/- adipo-/- double knockout mice. It will provide theoretical basis that regulating the function of proteins and protein therapy by dynamic post-translational modifications more accurately and more specifically, and provide promising target and method in treating CAD.

本课题前期研究发现：冠心病者冠状动脉形成粥样斑块，而同一大环境下同为中动脉的乳内动脉及桡动脉却不易发生，且病变冠脉外周心外膜脂肪内分泌功能紊乱，内质网应激，表明心脏局部的微环境影响冠脉病变的发生；在冠心病动物模型的心外膜脂肪局部转染脂联素基因（apM1）以抗动脉粥样硬化，效果欠佳；后续研究发现，心外膜脂肪在内质网应激状态下，脂联素翻译后修饰过程发生障碍，mRNA水平增加并未相应增加脂联素蛋白。据此，本课题以翻译后修饰的视角，从在体、亚细胞及动物模型三层面进一步探讨冠心病心脏局部微环境下脂联素糖基化的变化规律，利用SDS/PAGE法及质谱分析法精确分析脂联素各糖基化位点的变化；并在apoE-/- adipo-/- 双敲除小鼠中研究翻译后修饰各阶段脂联素形式抗粥样硬化效果及机制。为通过动态蛋白质翻译后修饰来更精细、专一的调控蛋白质功能及为蛋白质治疗提供理论依据，为冠心病治疗提供新靶点和新手段。

本课题前期研究发现：冠心病者冠状动脉形成粥样斑块，而同一大环境下同为中动脉的乳内动脉及桡动脉却不易发生，且病变冠脉外周心外膜脂肪内分泌功能紊乱，内质网应激，表明心脏局部的微环境影响冠脉病变的发生；在冠心病动物模型的心外膜脂肪局部转染脂联素基因（apM1）以抗动脉粥样硬化，效果欠佳；后续研究发现，心外膜脂肪在内质网应激状态下，脂联素翻译后修饰过程发生障碍，mRNA水平增加并未相应增加脂联素蛋白。据此，本课题以翻译后修饰的视角，从在体、亚细胞及动物模型三层面进一步探讨冠心病心脏局部微环境下脂联素糖基化的变化规律，利用SDS/PAGE法及质谱分析法精确分析脂联素各糖基化位点的变化；并在apoE-/- adipo-/- 双敲除小鼠中研究翻译后修饰各阶段脂联素形式抗粥样硬化效果及机制。为通过动态蛋白质翻译后修饰来更精细、专一的调控蛋白质功能及为蛋白质治疗提供理论依据，为冠心病治疗提供新靶点和新手段。