基于miR-124/CCL2信号通路探讨连翘酯苷A抑制内毒素炎症的作用机制研究

Endotoxin inflammation is an important pathological basis of critical illness, such as sepsis. In endotoxin inflammation, neutrophils and macrophages are important cells that produce inflammatory factors. Inhibiting excessive migration and aggregation of the neutrophils and macrophages is an effective means of attenuating endotoxin inflammation. Studies have shown that miRNAs-chemokines regulate neutrophils and macrophages in endotoxin inflammation. Our previous studies have found that Forsythoside A (FA), the main component of Forsythia suspensa, can significantly inhibit the migration and aggregation of neutrophils and macrophages, up-regulate the expression of miR-124, and decrease the expression of monocyte chemoattractant protein-1 (CCL2) mRNA. Therefore, we speculated that FA may inhibits the migration and aggregation of neutrophils and macrophages by regulating the miR-124/CCL2 pathway. Based on previous work, we will explore the effects of miR-124 and CCL2 on the migration and aggregation of neutrophils and macrophages and the molecular mechanism of FA by Morpholino gene knockdown, cell co-culture, qPCR and other techniques. This study propose new ideas for the study of anti-endotoxin inflammation mechanism of Chinese medicine, and provide experimental basis for the development and application of FA.

内毒素炎症是脓毒症等危重病的重要病理基础。在内毒素炎症中，中性粒与巨噬细胞是产生炎性因子的重要细胞，抑制上述炎细胞的过度迁移、聚集，是阻断内毒素炎症的有效手段。研究表明，miRNAs-趋化因子在内毒素炎症中对炎细胞具有重要的调控作用。本课题组研究发现，连翘酯苷A（Forsythoside A，FA）可显著抑制炎细胞的迁移、聚集，上调miR-124的表达，降低单核细胞趋化蛋白-1（CCL2） mRNA水平，故提出通过调控miR-124/CCL2通路抑制炎细胞炎症行为的FA抗内毒素炎症作用新机制。本项目以miR-124/CCL2信号通路为切入点，拟通过建立miR-124 knockdown模型，采用细胞共培养、qPCR等技术，探讨miR-124与CCL2对炎细胞炎症行为的影响及FA的干预作用，阐明FA抗内毒素炎症作用机制，为中药抗内毒素炎症机制研究提出新的思路，为FA的开发应用提供实验依据。

内毒素炎症是脓毒症等危重病的重要病理基础。在内毒素炎症中，中性粒与巨噬细胞是产生炎性因子的重要细胞，抑制上述炎细胞的过度迁移、聚集，是阻断内毒素炎症的有效手段。研究表明，miRNAs-趋化因子在内毒素炎症中对炎细胞具有重要的调控作用。. 本课题组前期研究发现，连翘酯苷A（Forsythoside A，FA）可显著抑制炎细胞的迁移、聚集，上调miR-124的表达，降低单核细胞趋化蛋白-1（CCL2）mRNA水平，故提出通过调控miR-124/CCL2通路抑制炎细胞炎症行为的FA抗内毒素炎症作用新机制。. 本项目主要研究了：（1）通过体内实验阐明：miR-124/CCL2对体内内毒素炎症调控及FA干预作用研；（2）：通过体外实验阐明：miR-124/CCL2对炎细胞行为调控及FA干预作用研究。实验结果显示：miR-124/CCL2信号在内毒素所致体内外炎症中均对巨噬细胞、中性粒细胞发挥了调控作用。通过上调miR-124，抑制CCL2的表达，可显著抑制巨噬细胞、中性粒细胞的迁移、聚集和炎症因子的释放。FA在体内外均可通过上调miR-124表达，抑制CCL2的释放，达到抑制巨噬细胞、中性粒细胞的迁移、聚集，发挥抗炎作用。. 本项目以miR-124/CCL2信号通路为切入点，探讨了miR-124与CCL2对炎细胞炎症行为的影响及FA的干预作用，阐明了FA抗内毒素炎症作用机制，为中药抗内毒素炎症机制研究提出新的思路，为FA的开发应用提供实验依据。