解毒化瘀颗粒通过miRNAs/NF-κB/ROS调控NLRP3炎症体活化拮抗肝衰竭的机制研究

Activation of Kupffer cells could lead Immune inflammatory damage,which was the pathological basis of hepatic failure occurrence and development.NLRP3 Inflammasome is a new-found inflammation regulatory mechanism.It plays a key role in inflammation initiation, and maintain.In previous work, the expression profile of miRNA/mRNA in the treatment of liver failure by Jiedu Huayu granule was constructed.We also found the different miRNAs function mainly  enrichment in signaling pathways which were related to NLRP3 inflammasomes activation,such as NF-κB,NADPH oxidase,Kupffer cells polarization.However,Its precise regulation relationship is not clear.Our project group put forward a research hypothesis"Jiedu Huayu granule treated of liver failure through miRNAs intervention Kupffer cell NF- kappa B, ROS signal pathway, inhibit the activation of NLRP3 Inflammasome".We would carry out depth research by using technology such as slow virus interference and some steps such as miRNAs optimization, target gene regulation, the upstream transcription regulation, verification in vivo and so on.there are two main aims: To find and identify miRNAs with the most control inflammatory activation of NLRP3;To clear Molecular mechanisms which is miRNAs after LPS stimulation kupffer cells precise regulation NLRP3 inflammation activation and intervention targets of Jieduhuayu Granule.

Kupffer细胞激活引发的免疫炎症损伤是肝衰竭发生、发展的病理学基础。NLRP3等炎症体是一种新发现的炎症调控机制，在炎症启动及维持上发挥关键性作用。项目组前期构建了解毒化瘀颗粒拮抗肝衰竭的miRNA/mRNA表达谱，并在kupffer细胞中验证，发现其疗效机制可能是通过调控miRNA，进而影响下游NF-κB表达、ROS生成及NLRP3炎症体活化等有关，其精确调控关系尚未明确；据此提出“解毒化瘀颗粒通过miRNAs干预kupffer细胞内NF-κB、ROS信号通路，抑制NLRP3炎症体活化，拮抗肝衰竭”的研究假说。将应用慢病毒干扰等技术，按miRNAs优选、下游靶基因调控、上游转录子调控及体内验证等步骤开展深入研究，旨在发现及鉴定最具调控NLRP3炎症体活化作用的miRNAs；明确miRNAs在LPS刺激kupffer细胞后精确调控NLRP3炎症体活化的分子机制及解毒化瘀颗粒的干预靶点。

Kupffer细胞激活引发的免疫炎症损伤是肝衰竭发生、发展的病理学基础。NLRP3炎症体是一种新发现的炎症调控机制，在炎症启动及维持上发挥关键性作用。项目围绕miRNAs/NF-κB/ROS调控NLRP3活化开展相关工作，利用网络药理学研究方法解析了解毒化瘀颗粒调控免疫炎症机制拮抗肝衰竭的网络药理机制；发现了解毒化瘀颗粒调控NF-κB/ROS相关信号途径减弱NLRP3炎症体活化抑制kupffer细胞炎性应激强度的分子机制，进一步明确了解毒化瘀颗粒调控miRNA-223-3P表达抑制NLRP3活化的相关机制途径，且构建了解毒化瘀颗粒调控miRNA-223-3P表达形成的多维机制网络。研究结果明确了解毒化瘀颗粒抑制NLRP3炎症体活化减轻肝衰竭炎性损伤的分子机制，为中医药防治肝衰竭提供了新思路、新方法，促进了肝衰竭中医“毒浊”理论的发展及相关方药的深入研发与应用。