基于FKN/Syk/p38MAPK信号通路研究淫羊藿苷抗动脉粥样硬化作用及microRNA调控机制

Atherosclerosis (AS) is a common disease which is harmful to human health, but its pathogenesis is not very clear. Studies have shown that combination of the chemokine fractalkine (FKN) released by the endothelial cell and its receptor CX3CR1 resulted in the injury of endothelial cells, and then initiated AS. But FKN can induce p38MAPK expression by spleen tyrosine kinase (Syk), through inflammatory mechanism involved in AS, but the regulation mechanism is not very clear. Icariin (ICA) is the mainly active ingredients of Traditional Chinese Medicine epimedium, preliminary study confirmed that ICA can occur through the development of p38MAPK signal transduction pathway of AS, according to the association of the upstream and downstream signaling proteins. In this study, FKN, Syk and p38MAPK signal transduction will to be cascaded, the effect and mechanism of ICA against AS are going to be studied. At the same time, the miRNAs expression profiles in thoracic aorta of ApoE-/- mouse will to be detected, the miRNAs related to the FKN/Syk/p38MAPK signaling pathways will to be found, the potential exact signaling pathway will to be constructed and verified by Real-time PCR and Western blot.

动脉粥样硬化（AS）是危害人类健康的常见疾病，但其发病机制尚不十分明确。趋化因子（FKN）能与其受体CX3CR1结合，导致内皮细胞损伤，从而启动AS发生，而FKN可介导脾酪氨酸激酶（Syk）诱导p38MAPK的表达，通过炎症机制参与AS的发生发展，但其调控机制尚不清楚。淫羊藿苷（ICA）为传统中药淫羊藿的主要药效成分，前期研究表明，ICA可通过p38MAPK信号通路干预AS发生发展，根据上下游信号通路蛋白的关联性，本研究拟将FKN、Syk、p38MAPK等信号通路蛋白级联，探讨ICA抗AS作用及其机制。同时应用miRNAs芯片检测ApoE-/-小鼠胸主动脉miRNAs表达谱，在微阵列数据中寻找差异表达的miRNA，将其调控的靶基因与FKN/Syk/p38MAPK信号通路进行关联，构建AS发病潜在的确切信号通路，并以Real-time PCR、Western blot技术验证。

动脉粥样硬化（AS）是危害人类健康的常见疾病，但其发病机制尚不十分明确。趋化因子（FKN）能与其受体CX3CR1结合，导致内皮细胞损伤，从而启动AS发生，而FKN可介导脾酪氨酸激酶（Syk）诱导p38MAPK的表达，通过炎症机制参与AS的发生发展，但其调控机制尚不清楚。淫羊藿苷（ICA）为传统中药淫羊藿的主要药效成分，前期研究表明，ICA可通过p38MAPK信号通路干预AS发生发展，根据上下游信号通路蛋白的关联性，本研究将FKN、Syk、p38MAPK等信号通路蛋白级联，探讨ICA抗AS作用及其机制。同时应用lncRNA及miRNAs芯片检测ApoE-/-小鼠胸主动脉lncRNA及miRNAs表达谱，在微阵列数据中寻找差异表达的lncRNA和miRNA，将其调控的靶基因与FKN/Syk/p38MAPK信号通路进行关联，构建AS发病潜在的确切信号通路。.本研究发现ICA通过FKN/Syk/p38MAPK信号通路发挥抗动脉粥样硬化作用。并分别从平滑肌细胞、单核细胞和内皮细胞角度出发，探讨ICA抗动脉粥样硬化的机制。.在平滑肌细胞方面，lncRNA H19通过调控PKC βⅠ介导了ICA抑制HA-VSMCs的增殖、迁移作用；并以miR-205-5p/ERBB4/AKT信号通路为切入点，发现miR-205-5p是ICA抑制ox-LDL诱导的HA-VSMCs增殖与迁移的分子靶点。.在单核细胞方面，ICA通过lncRNA 042398抑制ox-LDL诱导的RAW264.7细胞的增殖与泡沫化。并以miR-672-5p/AKT3/NF-κB信号通路为切入点，发现miR-672-5p可作为抑制ox-LDL诱导的RAW264.7泡沫化和炎症反应、治疗AS的分子靶点。.在内皮细胞方面，基于lncRNA H19/miR-148b-3p/ELF5通路，发现ICA抑制ox-LDL诱导的HUVECs内皮间质转化的作用是通过lncRNA H19/miR-148b-3p/ELF5信号通路实现的。并在体内研究证实，ICA对ApoE-/-小鼠AS的保护作用是通过lncRNA H19实现的，说明lncRNA H19是ICA抑制动脉粥样硬化的重要分子靶点。.本研究从非编码RNA的角度对于ICA的抗AS潜能作出了更为全面的阐释，为AS的诊断和治疗提供新的分子标靶和干预策略。