Lu Di, Xia Qiaoyun, Yang Zhiyu, Gao Shanjun, Sun Suofeng, Luo Xiaoying, Li Zhen, Zhang Xiulei, Han Shuangyin, Li Xiuling, Cao Mingbo
Department of Gastroenterology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, School of Clinical Medicine, Henan University, Zhengzhou, China.
Microbiome Laboratory, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, China.
Ann Transl Med. 2021 Apr;9(8):661. doi: 10.21037/atm-21-471.
ENO3 expression is upregulated in Non-alcoholic fatty liver disease (NAFLD) patient tissues, demonstrated that ENO3 might play crucial roles in NAFLD. However, the mechanism of ENO3 in NAFLD remains unclear. Therefore, this study aimed to investigate the regulatory mechanism of ENO3 in the progression of non-alcoholic steatohepatitis (NASH) and NASH model.
and NASH model were established by methionine-choline deficient (MCD)-diet feeding and high free fatty acid (HFFA) induction in L02 cells. Loss and gain function of ENO3 and GPX4 was performed to study the mechanism in NASH. Western blot was used to detect the expression of ENO3 and GPX4. Hematoxylin and eosin (H&E), picrosirius Red and Oil Red O staining was used to evaluate histopathology of liver in NASH model. Ferroptosis indicators were measured by assay kits according to the manufacturer's instructions.
NASH mouse model was successfully established induced by MCD diet with steatosis, inflammatory infiltration, ballooning and fibrosis observed in the liver tissue. The expression of ENO3 and GPX4 was significantly elevated while ferroptosis was inhibited in NASH mice and cell model. Upregulation of both ENO3 and GPX4 could promote the lipid accumulation in L02 cells. In addition, overexpressed ENO3 attenuated the status of ferroptosis.
In the present study, we demonstrate that ENO3 promoted the progression of NASH by negatively regulating ferroptosis via elevating GPX4 expression and lipid accumulation. These findings provided solid foundation for the mechanism of ferroptosis on the progression of NASH regulated by ENO3, suggesting that ENO3 may be a potential therapeutic target for NASH.
非酒精性脂肪性肝病（NAFLD）患者组织中ENO3表达上调，表明ENO3可能在NAFLD中起关键作用。然而，ENO3在NAFLD中的作用机制仍不清楚。因此，本研究旨在探讨ENO3在非酒精性脂肪性肝炎（NASH）进展及NASH模型中的调控机制。
通过蛋氨酸-胆碱缺乏（MCD）饮食喂养和高游离脂肪酸（HFFA）诱导在L02细胞中建立NASH模型。进行ENO3和GPX4的功能缺失和获得实验以研究NASH中的机制。采用蛋白质免疫印迹法检测ENO3和GPX4的表达。用苏木精-伊红（H&E）、天狼星红和油红O染色评估NASH模型中肝脏的组织病理学。根据制造商的说明使用检测试剂盒测量铁死亡指标。
通过MCD饮食成功建立了NASH小鼠模型，肝脏组织出现脂肪变性、炎症浸润、气球样变和纤维化。NASH小鼠和细胞模型中ENO3和GPX4的表达显著升高，而铁死亡受到抑制。ENO3和GPX4的上调均可促进L02细胞中的脂质积累。此外，过表达的ENO3减轻了铁死亡状态。
在本研究中，我们证明ENO3通过上调GPX4表达和脂质积累负向调节铁死亡，从而促进NASH的进展。这些发现为ENO对NASH进展的铁死亡机制提供了坚实的基础，表明ENO3可能是NASH的潜在治疗靶点。
