Zhu Hong, Zhang Mengda, Ye Ying, Liu Zhenni, Wang Jianpeng, Wu Xue, Lv Xiongwen
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Institute for Liver Diseases of Anhui Medical University, Hefei 230032, China.
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Institute for Liver Diseases of Anhui Medical University, Hefei 230032, China.
Biochem Pharmacol. 2023 Sep;215:115753. doi: 10.1016/j.bcp.2023.115753. Epub 2023 Aug 21.
Alcohol use is a major risk factor for death and disability, resulting in a significant global disease burden. Alcoholic steatohepatitis (ASH) reflects an acute exacerbation of alcoholic liver disease (ALD) and is a growing health care and economic burden worldwide. Pyroptosis plays a central role in the pathogenesis of ASH. Nt5e (CD73) is a cell surface ecto-5'-nucleotidase, which is a key enzyme that converts the proinflammatory signal ATP to the anti-inflammatory mediator adenosine (ADO). Studies have found that CD73 is involved in multiple diseases and can alleviate gasdermin D (GSDMD)-mediated pyroptosis; however, its role and mechanism in ASH are not explicit.
To investigate the role and mechanisms of CD73-mediated hepatocyte pyroptosis in alcohol-induced liver injury through in vivo and in vitro experiments.
CD73 knockout (CD73) mice, wild-type (WT) mice, and AML-12 cells were used to evaluate the effect of CD73 on hepatocyte pyroptosis in vivo and in vitro. A combination of molecular and histological methods was performed to assess pyroptosis and investigate the mechanism both in vivo and in vitro.
The protein expression of CD73 and pyroptosis pathway-associated genes was increased significantly in hepatocyte injury model both in vivo and in vitro. In vivo, CD73 knockout dramatically aggravated inflammatory damage, lipid accumulation, and hepatocyte pyroptosis in the liver. In vitro, overexpression of CD73 by pEGFP-C1/CD73 can decrease NLRP3 inflammasome activation and pyroptosis in hepatocytes. Further analysis revealed that the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway is a possible mechanism of CD73 regulation. Meanwhile, this pathological process was inhibited after the use of PI3K inhibitors.
Our results show a novel function of CD73 regulates hepatocytes pyroptosis and highlights the therapeutic opportunity for reducing the disease process in ALD.
饮酒是导致死亡和残疾的主要风险因素，在全球造成了重大的疾病负担。酒精性脂肪性肝炎（ASH）反映了酒精性肝病（ALD）的急性加重，在全球范围内是日益加重的医疗和经济负担。细胞焦亡在ASH的发病机制中起核心作用。Nt5e（CD73）是一种细胞表面外5'-核苷酸酶，是将促炎信号三磷酸腺苷（ATP）转化为抗炎介质腺苷（ADO）的关键酶。研究发现，CD73参与多种疾病，并可减轻gasdermin D（GSDMD）介导的细胞焦亡；然而，其在ASH中的作用和机制尚不清楚。
通过体内和体外实验研究CD73介导的肝细胞焦亡在酒精性肝损伤中的作用及机制。
利用CD73基因敲除（CD73）小鼠、野生型（WT）小鼠和AML-12细胞评估CD73在体内和体外对肝细胞焦亡的影响。采用分子和组织学方法相结合的方式评估细胞焦亡，并研究体内和体外的机制。
在体内和体外的肝细胞损伤模型中，CD73的蛋白表达以及与细胞焦亡途径相关的基因均显著增加。在体内，敲除CD73会显著加重肝脏的炎症损伤、脂质蓄积和肝细胞焦亡。在体外，通过pEGFP-C1/CD73过表达CD73可降低肝细胞中NLRP3炎性小体的激活和细胞焦亡。进一步分析表明，磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路是CD73调节的一种可能机制。同时，使用PI3K抑制剂后，这一病理过程受到抑制。
我们的研究结果显示了CD73调节肝细胞焦亡的新功能，并突出了减少ALD疾病进程的治疗机会。
