圣草酚调控UBA52表达对代谢相关脂肪性肝病的体内和体外作用

doi:10.11958/20252251

天津医药 ›› 2025, Vol. 53 ›› Issue (9): 916-922.doi: 10.11958/20252251

圣草酚调控UBA52表达对代谢相关脂肪性肝病的体内和体外作用

林义伟¹(), 魏谭军¹^,^△(), 陈飞¹, 肖成¹, 袁烈², 王毅¹

1 达州市中西医结合医院药学科（邮编635000）
2 重庆医科大学药学院

收稿日期:2025-06-13 修回日期:2025-07-09 出版日期:2025-09-15 发布日期:2025-09-16
通讯作者: ^△E-mail：404477285@qq.com
作者简介:林义伟（1988），男，主管药师，主要从事药物分析及药剂学方面研究。E-mail：lyw105835@163.com
基金资助:
四川省中央引导地方科技发展专项项目(2023ZYD0283)

The in vivo and in vitro effects of Eriodictyol on metabolic dysfunction-associated steatotic liver disease by regulating UBA52 expression

LIN Yiwei¹(), WEI Tanjun¹^,^△(), CHEN Fei¹, XIAO Cheng¹, YUAN Lie², WANG Yi¹

1 Department of Pharmacy, Dazhou Integrated TCM & Western Medicine Hospital, Dazhou 635000, China
2 College of Pharmacy, Chongqing Medical University

Received:2025-06-13 Revised:2025-07-09 Published:2025-09-15 Online:2025-09-16
Contact: ^△E-mail: 404477285@qq.com

林义伟, 魏谭军, 陈飞, 肖成, 袁烈, 王毅. 圣草酚调控UBA52表达对代谢相关脂肪性肝病的体内和体外作用[J]. 天津医药, 2025, 53(9): 916-922.

LIN Yiwei, WEI Tanjun, CHEN Fei, XIAO Cheng, YUAN Lie, WANG Yi. The in vivo and in vitro effects of Eriodictyol on metabolic dysfunction-associated steatotic liver disease by regulating UBA52 expression[J]. Tianjin Medical Journal, 2025, 53(9): 916-922.

摘要/Abstract

摘要：

目的基于体内和体外水平探究圣草酚（ERI）调控泛素核糖体融合蛋白52（UBA52）表达对代谢相关脂肪性肝病发展的影响。方法采用高脂饮食诱导建立小鼠代谢相关脂肪性肝病模型。将小鼠随机分为正常对照组（Normal组）、模型组（Model组）、低剂量ERI组（ERI-L组，50 mg/kg ERI）、高剂量ERI组（ERI-H组，100 mg/kg ERI），每组12只。油红O染色观察小鼠肝脏组织的脂质沉积；HE染色观察小鼠肝脏组织病理学变化；酶联免疫吸附试验（ELISA）法检测小鼠血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、低密度脂蛋白胆固醇（LDL-C）、总胆固醇（TC）和甘油三酯（TG）水平；免疫印迹法检测肝脏组织中UBA52蛋白表达。采用0.5 mmol/L油酸处理HepG2细胞，诱导体外代谢相关脂肪性肝病模型。将HepG2细胞随机分为对照组（Control组）、油酸诱导组、低浓度ERI组（50 μmol/L ERI）、高浓度ERI组（100 μmol/L ERI）、高浓度ERI+si-NC组（100 μmol/L ERI+转染si-NC）和高浓度ERI+si-UBA52组（100 μmol/L ERI+转染si-UBA52）。油红O染色检测各组HepG2细胞的脂质沉积；ELISA检测各组HepG2细胞中TG、TC和氧化应激相关因子超氧化物歧化酶（SOD）、丙二醛（MDA）水平；免疫印迹法检测HepG2细胞中UBA52、p62、LC3Ⅱ、LC3Ⅰ自噬相关蛋白表达。结果与Normal组相比，Model组血清ALT、AST、LDL-C、TC和TG水平及肝脏组织UBA52蛋白表达升高（P<0.05），肝脏脂质沉积增多，病理损伤严重，脂质沉积面积占比及非酒精性脂肪性肝病（NAFLD）活动度评分升高（P<0.05）；ERI-L组、ERI-H组小鼠相应指标变化低于Model组（P<0.05），且ERI-H组更低（P<0.05），肝脏脂质沉积减少，病理损伤减轻。与Control组相比，油酸诱导组HepG2细胞TG、TC、MDA水平、脂滴面积占比及UBA52蛋白表达升高，SOD水平、p62、LC3Ⅱ/LC3Ⅰ降低（P<0.05）；低浓度ERI组、高浓度ERI组对代谢相关脂肪性肝病有治疗作用（P<0.05），且敲低UBA52表达后加强了ERI对代谢相关脂肪性肝病的治疗作用。结论 ERI可能通过下调UBA52的表达在体内体外水平减缓代谢相关脂肪性肝病的进展。

关键词: 自噬, 氧化性应激, 代谢相关脂肪性肝病, 圣草酚, 泛素核糖体融合蛋白52

Abstract:

Objective To investigate the effect of Eriodictyol (ERI) on the development of metabolic dysfunction-associated steatotic liver disease by regulating the expression of ubiquitin A 52 (UBA52) at both in vivo and in vitro levels. Methods A mouse metabolic dysfunction-associated steatotic liver disease model was established using a high-fat diet induction. The mice were randomly separated into the normal control group (normal group), the model group, the low-dose ERI group (ERI-L group, 50 mg/kg ERI) and the high-dose ERI group (ERI-H group, 100 mg/kg ERI), with 12 mice in each group. Oil red O staining was applied to observe lipid deposition in mouse liver tissue. HE staining was applied to observe pathological changes in mouse liver tissue. ELISA method was applied to detect serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC) and triglycerides (TG) in mice. The expression of UBA52 protein in liver was detected by Western blot assay. HepG2 cells were treated with 0.5 mmol/L oleic acid to induce an in vitro metabolic dysfunction-associated steatotic liver disease model. HepG2 cells were randomly divide into the control group, the oleic acid induced group, the low concentration ERI group (ERI low group, 50 μmol/L ERI), the high concentration ERI group (ERI high group, 100 μmol/L ERI), the high concentration ERI+si-NC group (ERI high+si-NC group, 100 μmol/L ERI+transfected with si-NC) and the high concentration ERI+si-UBA52 group (ERI high+si-UBA52 group, 100 μmol/L ERI+transfected with si-UBA52). Oil red O staining was applied to detect lipid deposition in HepG2 cells of each group. ELISA method was applied to detect the levels of TG, TC, SOD and MDA in HepG2 cells in each group. Immunoblotting was used to detect the expression levels of UBA52, p62 and autophagy related proteins in HepG2 cells. Results Compared with the normal group, serum levels of ALT, AST, LDL-C, TC, TG and the expression of UBA52 protein in liver tissue were increased in the model group (P<0.05), and the lipid deposition in liver increased, pathological damage was severe, and the proportion of lipid deposition area and non-alcoholic fatty liver disease (NAFLD) activity score were also increased (P<0.05). Changes in the corresponding indicators in the ERI-L group and the ERI-H group were opposite to those of the model group (P<0.05), and the ERI-H group was even lower (P<0.05). The lipid deposition in liver decreased and the pathological damage was alleviated. Compared with the control group, the levels of TG, TC, MDA, the proportion of lipid droplet area and the expression of UBA52 protein were increased in HepG2 cells of the oleic acid-induced group, while the levels of SOD, p62 and LC3Ⅱ/LC3Ⅰ decreased (P<0.05). Changes in the corresponding indicators of the low-concentration ERI group and the high-concentration ERI group were opposite to those of the oleic acid-induced group (P<0.05), and the therapeutic effect of ERI on metabolic dysfuntion-associated steatotic liver disease was enhanced after knocking down the expression of UBA52. Conclusion ERI may slow down the progression of metabolic dysfuntion-associated steatotic liver disease by down-regulating the expression of UBA52 at both in vivo and in vitro levels.

Key words: autophagy, oxidative stress, metabolic dysfunction-associated steatotic liver disease, eriodictyol, ubiquitin A 52

中图分类号:

R575

图/表 8

图1 油红O染色观察小鼠肝脏组织脂质沉积（×200）

Fig.1 Lipid deposition in mouse liver observed by oil red O staining (×200)

图2 HE染色观察小鼠肝脏组织病理学变化（×200）

Fig.2 Histopathological changes of liver tissue in mice observed by HE staining (×200)

表1 各组小鼠血清ALT、AST、LDL-C、TC和TG水平的比较（n=12，$\bar{x}±s$）

Tab.1 Comparison of serum ALT, AST, LDL-C, TC and TG levels between the four group of mice

组别	ALT/（U/L）		AST/（U/L）
Normal组	22.38±2.17		104.72±8.82
Model组	79.58±6.20^a		158.12±12.64^a
ERI-L组	60.26±5.54^b		134.49±10.43^b
ERI-H组	34.69±3.05^bc		109.28±7.65^bc
F	380.091^＊＊		72.496^＊＊
组别	LDL-C/（mmol/L）	TC/（mmol/L）	TG/（mmol/L）
Normal组	0.31±0.03	3.04±0.22	0.96±0.07
Model组	0.98±0.10^a	6.32±0.56^a	1.72±0.15^a
ERI-L组	0.73±0.07^b	5.10±0.43^b	1.48±0.15^b
ERI-H组	0.45±0.05^bc	3.35±0.31^bc	1.10±0.10^bc
F	233.158^＊＊	177.107^＊＊	97.095^＊＊

图3 免疫印迹法检测小鼠肝脏组织中UBA52蛋白表达 A：Normal组；B：Model组；C：ERI-L组；D：ERI-H组。

Fig.3 The expression of UBA52 protein in mouse liver tissue detected by Western blot assay

图4 油红O染色观察HepG2细胞脂质沉积（×400）

Fig.4 Lipid deposition in HepG2 cells observed by oil red O staining (×400)

表2 各组HepG2细胞中TG、TC和SOD、MDA水平的比较（n=6，$\bar{x}±s$）

Tab.2 Comparison of TG, TC, SOD and MDA levels between six groups of HepG2 cells

组别	TG（mmol/g）	TC（mmol/g）	SOD（U/mg）	MDA（μmol/g）
Control组	0.12±0.01	0.18±0.02	52.65±4.27	1.02±0.08
油酸诱导组	0.40±0.04^a	0.43±0.04^a	16.15±1.86^a	2.84±0.20^a
低浓度ERI组	0.29±0.03^b	0.35±0.03^b	27.22±2.36^b	2.38±0.20^b
高浓度ERI组	0.19±0.01^bc	0.27±0.02^bc	38.49±3.50^bc	1.71±0.17^bc
高浓度ERI+ si-NC组	0.19±0.02	0.26±0.03	39.02±3.88	1.69±0.17
高浓度ERI+ si-UBA52组	0.13±0.01^de	0.19±0.02^de	50.31±4.75^de	1.24±0.12^de
F	128.700^＊＊	72.000^＊＊	89.447^＊＊	107.098^＊＊

图5 免疫印迹法检测HepG2细胞中UBA52、p62、LC3Ⅰ、LC3Ⅱ蛋白表达 A：Control组；B：油酸诱导组；C：低浓度ERI组；D：高浓度ERI组；E：高浓度ERI+si-NC组；F：高浓度ERI+si-UBA52组。

Fig.5 The expression levels of UBA52, p62, LC3Ⅰ and LC3Ⅱ proteins in HepG2 cells detected by Western blot assay

表3 各组HepG2细胞中UBA52、p62、LC3Ⅰ、LC3Ⅱ蛋白表达水平比较（n=6，$\bar{x}±s$）

Tab.3 Comparison of UBA52, p62, LC3Ⅰ and LC3Ⅱ proteins between six groups of HepG2 cells

组别	UBA52	p62	LC3Ⅱ/LC3Ⅰ
Control组	0.20±0.02	0.93±0.09	0.89±0.09
油酸诱导组	0.86±0.09^a	0.23±0.02^a	0.26±0.03^a
低浓度ERI组	0.70±0.07^b	0.42±0.04^b	0.46±0.05^b
高浓度ERI组	0.48±0.05^bc	0.63±0.06^bc	0.68±0.07^bc
高浓度ERI+si-NC组	0.47±0.05	0.62±0.06	0.70±0.07
高浓度ERI+si-UBA52组	0.25±0.03^de	0.82±0.08^de	0.86±0.09^de
F	120.547^＊＊	99.489^＊＊	71.727^＊＊

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圣草酚调控UBA52表达对代谢相关脂肪性肝病的体内和体外作用

The in vivo and in vitro effects of Eriodictyol on metabolic dysfunction-associated steatotic liver disease by regulating UBA52 expression

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