基于miR-23b/ELAVL1通路研究茵陈蒿汤协同ucMSCs-exosomes调控急性肝衰竭肝细胞焦亡的效应机制

Acute liver failure(ALF) is one of the important and difficult problems in the liver disease, and the therapeutic effect of simple western medicine is poor. The pyroptosis of hepatocytes is the main factor in accelerating the process of ALF. The combination of tranditional Chinese medicine prescription of Yinchenhao Decoction and western medicine in the treatment of liver failure has showed exact clinical curative effect. Our earlier studies have showed synergistic therapeutic effect of Yinchenhao decoction and umbilical cord mesenchymal stem cells (ucMSCs) on acute liver failure, and miR-23b expression was significantly increased in ucMSCs-exosomes treated with Yinchenhao Decoction. This project will investigate the mechanism of Yinchenhao decoction in regulating the expression of miR-23b in ucMSCs-exosomes and the mechanism of miR-23b in inhibiting the pyroptosis of hepatocytes. The project will explore the mechanism of the synergistic therapeutic effect of Yinchenhao decoction and ucMSCs on ALF from a new perspective and provide therotical and experimental evidence for the treatment of ALF with Traditional Chinese Medicine and Western medicine.

急性肝衰竭是肝病防治领域中的重点与难点问题之一，单纯西医治疗效果不理想。研究表明肝细胞焦亡是导致急性肝衰竭进程加速的重要因素。中医经方茵陈蒿汤联合西医治疗肝衰竭已显示确切临床疗效。课题组前期研究发现茵陈蒿汤可协同脐带间充质干细胞（ucMSCs）调控急性肝衰竭肝细胞焦亡，且miR-23b的表达在茵陈蒿汤干预的ucMSCs-exosomes中明显增高。本项目拟以肝细胞焦亡为切入点，提出茵陈蒿汤可通过促进ucMSCs-exosomes中miR-23b含量的增高而抑制肝细胞焦亡和肝衰竭。因此，本项目拟在前期研究基础上建立急性肝衰体内外模型，探索茵陈蒿汤调控ucMSCs-exosomes中miR-23b的表达及miR-23b抑制肝细胞焦亡的作用途径及具体分子机制，进一步阐明茵陈蒿汤协同ucMSCs-exosomes调控肝衰的分子机理，为两者在肝衰中的联用提供理论基础和实验依据。

肝细胞焦亡是急性肝衰竭发生发展过程中的重要因素，因此本课题通过LPS联合D-Gal构建小鼠肝衰模型以及ATP联合LPS构建LO2细胞焦亡模型，探究茵陈蒿汤协同ucMSCs-exosomes治疗急性肝衰竭中肝细胞焦亡的作用机制研究。首先本研究拟通过体内实验，茵陈蒿汤联合或不联合ucMSCs-exosomes分组干预，检测肝酶表达情况、肝细胞焦亡以及相关蛋白的表达情况。通过体外实验，利用miRNA-Seq测序和生信富集分析等方法，探讨ucMSCs-exosomes在抑制肝细胞焦亡过程中具体的作用机制。通过本课题的研究，我们发现茵陈蒿汤联合ucMSCs-exosomes可以有效的抑制急性肝衰竭中肝细胞焦亡的发生，并且降低IL-1β和IL-18炎症细胞因子的释放。同时测序以及生信分析结果提示其中外泌体所释放的miR-423-5p在抑制肝细胞焦亡中发挥主导作用，而ZBP1则作为miR-423-5p的直接下游靶基因。本研究为探索ucMSCs调控急性肝衰竭中肝细胞焦亡奠定了坚实的工作基础。同时对于中西药结合应用于急性肝衰竭治疗提供了重要的启示性线索。在项目执行期间，撰写论文3篇，其中SCI发表1篇，中文统计源期刊已录用1篇，会议收录1篇。申请国家发明专利2项。培养硕士研究生3名，其中3名均为在读研究生。