Keratin 8重塑代谢稳态调控慢加急性肝衰竭肝细胞代偿性再生的作用及机制研究

成熟肝细胞代偿性增殖障碍是慢加急性肝衰竭疾病预后不良的关键原因，其中慢性肝病下肝细胞葡萄糖氧化磷酸化抑制，中间产物偏向氨基酸代谢通路是影响成熟肝细胞增殖激活的特征代谢异常模式。肝脏富含的角蛋白（Keratin，K）是骨架蛋白中间丝家族最主要成员，申请人前期研究证实K8的R341H突变是慢加急性肝衰竭的独立危险因素，预实验发现R341H突变引起K8的280位丝氨酸（S280）磷酸化下调介导了肝细胞葡萄糖氧化代谢和增殖显著抑制，结合K8参与肝癌增殖以及多组织的衰竭保护和再生修复功能，由此推测调控K8的S280磷酸化可靶向促进代谢稳态恢复而重建肝细胞增殖能力，促进肝再生。本研究拟通过细胞实验、临床标本及已有的敲除小鼠，采用qPCR、蛋白印迹、免疫共沉淀、免疫荧光等技术阐明K8调控肝再生的作用及机制，有望为慢加急性肝衰竭治疗提供新靶点。

Compensatory proliferation of mature hepatocytes has been identified as one of the key reasons for the poor prognosis of acute-on-chronic liver failure. The inhibition of glucose oxidative phosphorylation of hepatocytes in chronic liver disease due to the metabolic intermediates biased amino acid metabolism pathway are the characteristic metabolic abnormalities that hamper the proliferation abilities of mature hepatocytes. Keratin (K), as the most important member of the intermediate filament family of cytoskeletal proteins. The applicant’s previous studies have confirmed that the R341H mutation of K8 is an independent risk factor for the development of chronic acute liver failure. Preliminary experiments have found that R341H mutation causes K8 down-regulation of Serine (S280) phosphorylation that mediates significant inhibition of hepatocyte glucose oxidation and proliferation ,and as the literatures reported that K8 was involved in liver cancer proliferation and multi-organ failure protection and regenerations. Therefore, it is speculated that the regulation of K8 S280 phosphorylation may be another novel target to restore the metabolic homeostasis of hepatocytes and promote liver regeneration. This study intends to clarify the role and mechanism of K8 in regulating liver regeneration via using qPCR, western blotting, immunoprecipitation, immunofluorescence and other techniques in vitro experiments, clinical specimens and k8 knockout mice. This program may help to identify the mechanism of K8 in regulating liver regeneration, which is expected to provide new targets for the treatments of acute-on-chronic acute liver failure.