肝衰竭是发病机制复杂，死亡率高的危重病，病理表现为肝细胞坏死，抑制肝细胞坏死是防治肝衰竭的重点与难点。项目组前期研究发现三黄茵赤汤具有防治肝衰竭的作用，病理显示肝细胞具有凋亡和程序性坏死（necroptosis）的特征，且RIP3显著升高。因此，我们推测肝衰竭发生了RIP3依赖的necroptosis。拟用正常和RIP3基因敲除小鼠建立急性、慢加急性肝衰竭模型，通过CRISPR/Cas9系统技术过表达和沉默RIP3基因建立肝细胞损伤模型。体内、体外正反两个角度验证RIP3及相关蛋白在肝细胞necroptosis中的作用。免疫荧光、免疫印迹和qPCR等技术检测caspase3-RIP3-CaMKII通路相关蛋白及mRNA的表达。一方面揭示防治肝衰竭的分子机制，另一方面寻找防治肝衰竭的新靶点。同时，进一步诠释三黄茵赤汤多途径、多靶点防治急性肝衰竭的科学内涵，为临床应用提供理论依据。

Inhibition of Liver cell necrosis is the key and difficult point for prevention and treatment of liver failure. Previous clinical and experimental Liver failure is a complex pathogenesis, high mortality of critically ill, pathological manifestations of liver cell necrosis, inhibition of liver cell necrosis prevention and treatment of liver failure is important and difficult. Early research found SHYCT with function of prevention and treatment of liver failure, meanwhile, found hepatocytes apoptosis and necroptosis pathological features, and a significant rise in RIP3. Therefore, we surmise that liver failure happened necroptosis of RIP3 dependent. We established Acute liver failure(ALF) and Acute on chronic liver failure(ACLF) animal model by RIP3 knockout mice and normal mice,and cell injury model by CRISPR / Cas9 system overexpression and silence of RIP3 gene in normal liver cell line. In vivo and in vitro from both positive and negative angles verification RIP3 and related proteins in the liver cells necroptosis. Use immunofluorescence , western blot and qPCR to detection caspase3- RIP3 - CaMKII pathways related protein and mRNA expression. On the one hand, revealed the molecular mechanism of prevention and treatment of hepatic failure, on the other hand, to find new targets for the prevention and treatment of liver failure. At the same time ,further interpretation of that decoction multi-way,multi-target prevention and treatment of liver failure, the scientific connotation, provide sufficient theoretical basis for clinical application.