基于MAPK与NOS信号crosstalk调控的艳山姜挥发油保护血管内皮损伤的机制研究

The cardiovascular diseases, leading cause of mortality and morbidity, always seriously damage to human being health. The prevention and therapeutics, as well as pathological mechanism have been highlighted by many researchers in clinical and basic medical field. At present, it is widely accepted that focus on protecting integrity of structure and function for arterial endothelium, and looking for novel drug targets to prevent against endothelial injury, because the endothelial injury is not only the key pathological process but also the initiating factor in the development and deterioration of cardiovascular diseases. Accumulating evidence indicated that the MAPK and NOS signals are highly involved in endothelial injury. However, there is few research on the crosstalk and/or common signaling pathway in the process of endothelial injury. The essential oil from Fructus A. zerumbet (Yanshanjiang, Guizhou province local characteristic Miao folk herbs), a bioactive ingredient, could protect against the endothelial injury induced by ox-LDL, and the mechanism maybe related to regulating the NOS-NO signal upon our previous research. The present study is design to provide the novel theoretical guidance and experimental basis for cardiovascular diseases mainly on the endothelial injury. We concentrated on clarifying the crosstalk and/or common signaling pathway between MAPK and NOS signals in endothelial cell injury induced by ox-LDL. Furthermore, we elucidated the protective effects of key targets of essential oil from Fructus A. zerumbet, and relevant mechanism, would-be the experimental basis and theoretical guidance for exploring the novel pharmacological effects and promoting modern research of ethno-drug, and for traditional folk medicine in-depth research and development.

心血管系统疾病是人类健康的头号杀手，其发病机制及防治成为当前医药工作者高度关注的热点问题。血管内皮损伤是心血管系统疾患发展与恶化的关键环节与始动因素，保护动脉内皮结构与功能的完整性，寻找其损伤的关键靶位及治疗药物成为当前研究的焦点。研究提示，MAPK与NOS是目前内皮细胞损伤的重要信号系统，但两者之间交互作用的研究迄今未见相关报道。本实验室前期研究证实，贵州地产特色民族药材--艳山姜的挥发油对ox-LDL诱导血管内皮细胞损伤具有显著的保护作用，其作用与调控NOS系统相关。本项目以艳山姜挥发油为研究对象，建立ox-LDL诱导内皮细胞损伤的模型，系统分析MAPK与NOS信号之间的交互作用和/或共同通路，为基于内皮损伤的心血管系统疾患的防治提供新的实验基础与理论指导，并进一步探讨艳山姜挥发油防治内皮损伤的作用及作用机制，发掘传统民族药新的药理作用，促进民族药现代化深层次研究开发。

心血管系统疾病是人类健康的头号杀手，其发病机制及防治成为当前医药工作者高度关注的热点问题。血管内皮损伤是心血管系统疾患发展与恶化的关键环节与始动因素，而大量资料提示内皮功能异常与丝裂原活化蛋白激酶（MAPK）与一氧化氮合酶（NOS）密切相关，但是MAPKs与NOS之间是否存在crosstalk有必要进行系统研究。我们的前期研究证实，艳山姜挥发油对氧化低密度脂蛋白（ox-LDL） 诱导血管内皮细胞损伤具有显著的保护作用，其作用与调控NOS-NO 系统及改善细胞凋亡密切相关。本课题以体外培养血管内皮细胞为研究对象，以动脉粥样硬化发病重要因素——ox-LDL复制血管内皮损伤模型，系统研究艳山姜挥发油（EOFAZ）对ox-LDL诱导损伤的人主动脉内皮细胞（HAECs）损伤的保护作用及对MAPKs与NOS信号调控。研究结果证实EOFAZ对ox-LDL诱导HAECs损伤具有保护作用，能促进细胞损伤的迁移和修复，其保护作用机制与抑制MAPK信号p38MAPK、JNK1/2/3、ERK1/2磷酸化以及调控eNOS和iNOS的平衡有关。ox-LDL诱导HAECs损伤过程中存在MAPK信号与NOS信号crosstalk，为临床血管功能损伤疾患的防治提供新的理论与实验基础。此外，初步探索艳山姜挥发油对LPS及高糖诱导血管内皮细胞损伤的机制，阐明EOFAZ对NF-κB活化与失活、核移位的影响；建立艳山姜挥发油（EOFAZ）的气相色谱-质谱（GC-MS）的特征指纹图谱，明确EOFAZ的抗炎、镇痛和内皮细胞保护作用的药效物质基础。本研究采用现代医学与生物学的研究方法与手段，为基于内皮损伤的心血管系统疾患的防治提供新的实验基础与理论指导；为发掘传统民族药新的药理作用，促进民族药现代化研究及其深层次研究开发提供实验基础。项目执行期间发表标注本项目资助的研究论文15篇，其中SCI论文5篇，核心期刊论文7篇。培养博士研究生1名，硕士研究生生6名。研究成果“艳山姜等四种贵州地产是药材生物活性与机制研究”获得贵州省科技进步三等奖，项目负责人排名第一；“基于血管内皮损伤关键靶位的药物新技术与新方法研究”获得贵州省高等学校科学研究优秀成果奖（自然科学奖）三等奖。