基于斑马鱼模型研究异甘草酸镁对酒精性肝损伤的保护作用机制

Alcoholic liver disease has become the second major cause of liver injury after viral hepatitis in China recently, which impairs people’s health seriously. However, the pathogenesis and therapeutic of acute alcoholic liver injury induced by binge drinking still remain to be further studied. In our early study, we took the lead in establishing the alcohol-induced liver injury model in zebrafish larvae and drug screening research have been carried on the basis of this model. We found that Magnesium Isoglycyrrhizinate（MgIG）, a drug independently developed in China with complete intellectual property rights, has liver protection effect on alcohol-induced liver injury. Further study revealed that paqr3b（PAQR3）may mediate the liver protection function of MgIG by zebrafish gene chip. Based on this, this study intends to construct paqr3b mutant and transgenic zebrafish by morpholine antisense oligonucleotide, CRISPR-Cas9 and transgenic technology. On the other hand, the expression silencing and overexpression cell lines of PAQR3 were constructed by using siRNA and overexpression vector at the cellular level. The pathogenesis of acute alcoholic liver injury and the liver protection mechanism of MgIG were elucidated from the perspective of PAQR3 in vivo and in vitro using molecular biology, cell biology and histology methods. The data of this study would help us to illuminate the pathogenesis of alcoholic liver injury and provide a reliable preclinical basis of the application of MgIG in the treatment of alcoholic-liver injury. The above contents have not been reported after retrieval.

近年来酒精性肝病已成为我国继病毒性肝炎后导致肝脏损伤的第二大病因，严重危害人民健康。但是酗酒引起肝脏急性损伤的机制尚不完全清楚，更缺乏有效的临床治疗措施。本课题组前期利用斑马鱼模式生物在国内率先建立了酒精诱导的急性酒精性肝损伤模型，在此基础上首次发现我国自主研发、拥有完全知识产权的药物---异甘草酸镁具有肝脏保护作用。通过斑马鱼基因芯片筛选分析发现paqr3b（PAQR3）可能介导了异甘草酸镁的肝脏保护功能。基于此，本研究拟应用CRISPR-Cas9以及转基因技术构建突变体/转基因斑马鱼，在细胞水平通过siRNA、过表达载体等技术构建表达沉默/过表达细胞株，继而以分子生物学、细胞生物学、组织学等方法从体内体外层面、从PAQR3这个全新的视角阐明酒精性肝损伤的发病机制及异甘草酸镁的保肝作用机制，为异甘草酸镁应用于酒精性肝损伤治疗提供可靠的临床前实验依据。以上内容经检索无报道。

饮酒（有害使用酒精）是加重全球疾病负担的一个重要因素，并被列为世界上导致早亡和残疾的第三大风险因素。随着我国经济和生活水平的不断提高，酗酒在我国呈现不断攀升的趋势，酒精性肝病的发病率也逐年升高。目前酒精已成为我国继病毒性肝炎后导致肝脏损伤的重要病因，严重危害人民健康。但是酗酒引起肝脏急性损伤的机制尚不完全清楚，更缺乏有效的临床治疗措施。因此，本课题着力于深入探讨酗酒引起的肝脏急性损伤的机制以及研发肝脏保护性药物，这具有重要的现实意义。研究结果提示：1）成功创建了急性酒精性肝损伤体内及体外模型；2）证实异甘草酸镁（MgIG）对急性酒精性肝损伤的疗效；3）异甘草酸镁通过调控脂质代谢、内质网应激发挥肝脏保护作用；4）PAQR3/SREBP2/HMGCR可能介导了异甘草酸镁的保肝作用。以上科研结果进一步阐明了异甘草酸镁对酒精性肝损伤的保护作用及其初步作用机制，为预防和治疗酒精性肝损伤提供可靠的临床前依据。