表儿茶素对对乙酰氨基酚诱导小鼠肝损伤的改善作用及其机制

doi:10.13481/j.1671-587X.20250606

吉林大学学报(医学版) ›› 2025, Vol. 51 ›› Issue (6): 1498-1507.doi: 10.13481/j.1671-587X.20250606

• 基础研究 • 上一篇

表儿茶素对对乙酰氨基酚诱导小鼠肝损伤的改善作用及其机制

于蕙源^1,²,金令³,于颖⁴,王雪⁵,王冰⁶()

^1.吉林大学第一医院放疗科，吉林长春 130021
^2.吉林大学动物医学学院临床兽医学系，吉林长春 130062
^3.吉林大学第一医院检验科，吉林长春 130021
^4.吉林大学第一医院生殖中心产前诊断中心，吉林长春 130021
^5.吉林省松原市吉林油田医院检验科，吉林松原 138000
^6.吉林省肿瘤医院放疗科，吉林长春 130012

收稿日期:2024-12-09 接受日期:2025-01-16 出版日期:2025-11-28 发布日期:2025-12-15
通讯作者: 王冰 E-mail:zisu8688@163.com
作者简介:于蕙源（1998-），男，吉林省长春市人，在读硕士研究生，主要从事放射肿瘤学方面的研究。
基金资助:
吉林省教育厅科研重点项目(JJKH20241337KJ)

Improvement effect of epicatechin on liver injury in mice induced by acetaminophen and its mechanism

Huiyuan YU^1,²,Ling JIN³,Ying YU⁴,Xue WANG⁵,Bing WANG⁶()

^1.Department of Radiotherapy，First Hospital，Jilin University，Changchun 130021，China
^2.Department of Clinical Veterinary Medicine，School of Veterinary Medicine，Jilin University，Changchun 130062，China
^3.Department of Clinical Laboratory，First Hospital，Jilin University，Changchun 130021，China
^4.Reproductive Center，Prenatal Diagnosis Center，First Hospital，Jilin University，Changchun 130021，China
^5.Department of Clinical Laboratory，Songyuan Jilin Oilfield Hospital，Jilin Province，Songyuan 138000，China
^6.Department of Radiotherapy，Tumor Hospital，Jilin Province，Changchun 130012，China

Received:2024-12-09 Accepted:2025-01-16 Online:2025-11-28 Published:2025-12-15
Contact: Bing WANG E-mail:zisu8688@163.com

摘要/Abstract

摘要：

目的探讨表儿茶素（EC）对对乙酰氨基酚（APAP）诱导小鼠肝损伤的改善作用，并阐明其可能的作用机制。方法将60只C57BL/6J小鼠随机分为空白对照组、APAP模型组、低剂量（10 mg·kg^-1）EC组、中剂量（20 mg·kg^-1）EC组和高剂量（40 mg·kg^-1）EC组，每组12只。除空白对照组外，其余各组小鼠给予腹腔注射APAP（200 mg·kg^-1）诱导肝损伤模型。APAP注射前1 h，低、中和高剂量EC组小鼠腹腔分别注射10、20和40 mg·kg^-1 EC。36只核因子E2相关因子2（Nrf2）缺陷小鼠（Nrf2^-/-小鼠）随机分为对照组、APAP组和APAP+EC组，每组12只。造模24 h后处死小鼠，收集小鼠血液和肝组织用于后续检测。采用HE染色观察各组小鼠肝组织病理形态表现，试剂盒检测各组小鼠血清中天冬氨酸氨基转移酶（AST）和丙氨酸氨基转移酶（ALT）活性及小鼠肝组织中髓过氧化物酶（MPO）活性和肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、丙二醛（MDA）、三磷酸腺苷（ATP）、谷胱甘肽（GSH）及亚铁离子（Fe2?）水平，Western blotting法检测各组小鼠肝组织中核因子κB（NF-κB）和Nrf2信号通路相关蛋白表达水平。结果 HE染色，与APAP模型组比较，不同剂量EC组APAP诱导小鼠肝病理损伤明显改善。与空白对照组比较，APAP模型组小鼠血清中ALT和AST活性明显升高（P<0.01）；与APAP模型组比较，低、中和高剂量EC组小鼠血清中ALT及AST活性明显降低（P<0.05或P<0.01）。与空白对照组比较，APAP模型组小鼠肝组织中MPO活性及TNF-α和IL-1β水平明显升高（P<0.01）；与APAP模型组比较，低、中和高剂量EC组小鼠肝组织中MPO活性及TNF-α和IL-1β水平明显降低（P<0.05或P<0.01）。与空白对照组比较，APAP模型组小鼠肝组织中MDA和Fe²⁺水平明显升高（P<0.05），ATP和GSH水平明显降低（P<0.05）；与APAP模型组比较，低、中和高剂量小鼠肝组织中MDA和Fe²⁺水平明显降低（P<0.05或P<0.01），ATP和GSH水平明显升高（P<0.05或P<0.01）。与空白对照组比较，APAP模型组小鼠肝组织中氨基酸交换转运蛋白（xCT）和谷胱甘肽过氧化物酶4（GPX4）蛋白表达水平明显降低（P<0.05）；与APAP模型组比较，低、中和高剂量小鼠肝组织中xCT和GPX4蛋白表达水平明显升高（P<0.01）。与空白对照组比较，APAP模型组小鼠肝组织中核因子κB（NF-κB） p-p65和磷酸化NF-κB抑制剂α（p-IκBα）蛋白表达水平明显升高（P<0.05）；与APAP模型组比较，低、中和高剂量EC组小鼠肝组织中NF-κB p-p65和p-IκBα蛋白表达水平明显降低（P<0.01）。与空白对照组比较，APAP模型组小鼠肝组织中Nrf2和血清素加氧酶1（HO-1）蛋白水平明显升高（P<0.05）；与APAP模型组比较，低、中和高剂量EC组小鼠肝组织中Nrf2和HO-1蛋白水平明显升高（P<0.01）。与对照组比较，APAP组Nrf2^-/-小鼠血清中AST和ALT活性及肝组织中MDA和Fe²⁺水平明显升高（P<0.01），肝组织中ATP和GSH水平明显降低（P<0.01）。结论 EC对APAP诱导的小鼠肝损伤具有改善作用，其机制可能与EC激活Nrf2/GPX4信号通路抑制铁死亡有关。

关键词: 表儿茶素, 对乙酰氨基酚, 肝损伤, 铁死亡, 核因子E2相关因子2

Abstract:

Objective To discuss the improvement effect of epicatechin（EC） on acetaminophen（APAP）- induced liver injury in the mice， and to clarify its possible mechanism. Methods A total of 60 C57BL/6J mice were randomly divided into blank control group， APAP model group， low dose of EC group （10 mg·kg^-1）， middle dose of EC group （20 mg·kg^-1） and high dose of EC group （40 mg·kg?1）， with 12 mice in each group. Except for blank control group， the mice in the other groups were given intraperitoneal injection of APAP （200 mg·kg?1） to establish the liver injury models. At 1 h before APAP injection， the mice in low， middle and high doses of EC groups were intraperitoneally injected with 10， 20 and 40 mg·kg^-1 EC， respectively. A total of 36 nuclear factor E2-related factor 2 （Nrf2） deficient mice （Nrf2^-/- mice） were randomly divided into control group， APAP group and APAP+EC group， with 12 mice in each group. After modeling 24 h， the mice were sacrificed， and the blood and liver tissue of the mice were collected for subsequent detection. HE staining was used to observe the pathomorphology of the liver tissue in the mice in various groups； kit assay was used to detect the activities of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in serum of the mice in various groups and the myeloperoxidase （MPO） activity and the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， malondialdehyde （MDA）， adenosine triphosphate （ATP）， glutathione （GSH） and ferrous ion （Fe2?） in liver tissue of the mice in various groups； Western blotting method was used to detect the expression levels of nuclear factor-κB （NF-κB） and Nrf2 signaling pathway-related proteins in liver tissue of the mice in various groups. Results The HE staining results showed that compared with APAP model group， the APAP-induced liver pathology injury in the mice in different doses of EC groups was significantly improved. Compared with blank control group， the levels of ALT and AST in serum of the mice in APAP model group were significantly increased （P<0.01）. Compared with APAP model group， the activities of ALT and AST in serum of the mice in low， middle and high doses of EC groups were significantly decreased （P<0.05 or P<0.01）. Compared with blank control group， the MPO activity and the levels of TNF-α and IL-1β in liver tissue of the mice in APAP model group were significantly increased （P<0.01）. Compared with APAP model group， the MPO activity and the levels of TNF-α and IL-1β in liver tissue of the mice in low， middle and high doses of EC groups were decreased （P<0.05 or P<0.01）. Compared with blank control group， the levels of MDA and Fe2? in liver tissue of the mice in APAP model group were significantly increased （P<0.05）， and the levels of ATP and GSH were significantly decreased （P<0.05）. Compared with APAP model group， the levels of MDA and Fe2? in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.05 or P<0.01）， and the levels of ATP and GSH were significantly increased （P<0.01）. Compared with blank control group， the expression levels of amino acid exchange transporter （xCT） and glutathione peroxidase 4 （GPX4） proteins in liver tissue of the mice in APAP model group were significantly decreased （P<0.05）； compared with APAP model group， the expression levels of xCT and GPX4 proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.01）. Compared with blank control group， the expression levels of nuclear factor-κB （NF-κB） p-p65 and phosphorylated NF-κB inhibitor α （p-IκBα） proteins in liver tissue of the mice in APAP model group were significantly increased （P<0.05）； compared with APAP model group， the expression levels of NF-κB p-p65 and p-IκBα proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly decreased （P<0.01）. Compared with blank control group， the expression levels of Nrf2 and heme oxygenase-1 （HO-1） proteins in liver tissue of the mice in APAP model group were significantly increased （P<0.05）； compared with APAP model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.01）. Compared with control group， the ALT level in serum and the levels of MDA and Fe²⁺ in liver tissue of the Nrf2^-/- mice in APAP group were significantly increased （P<0.01）， and the levels of ATP and GSH in liver tissue were significantly decreased （P<0.01）. Conclusion EC can improve APAP-induced liver injury in the mice， and its mechanism may be related to the inhibition of ferroptosis by activating the Nrf2/GPX4 signaling pathway.

Key words: Epicatechin, Acetaminophen, Liver injury, Ferroptosis, Nuclear factor E2-related factor 2

中图分类号:

R575

于蕙源,金令,于颖,王雪,王冰. 表儿茶素对对乙酰氨基酚诱导小鼠肝损伤的改善作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(6): 1498-1507.

Huiyuan YU,Ling JIN,Ying YU,Xue WANG,Bing WANG. Improvement effect of epicatechin on liver injury in mice induced by acetaminophen and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1498-1507.

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Group	ALT	AST
Blank control	32.75±17.52	47.12±22.56
APAP model	582.63±36.32^*	509.22±15.97^*
EC
Low dose	425.33±21.47^△	396.55±10.29^△
Middle dose	215.69±19.59^△△	189.96±30.21^△△
High dose	153.92±24.91^△△	109.85±26.91^△△

Group	MPO activity[λ_B/（U·L^-1）]	TNF-α level [ω_B/（ng·g^-1）]	IL-1β level [ω_B/（ng·g^-1）]
Blank control	1.023±0.352	119.36±10.32	47.12±22.56
APAP model	5.336±1.096^*	756.83±63.98^*	509.22±15.97^*
EC
Low dose	4.589±0.966^△	572.41±98.25^△	396.55±10.29^△
Middle dose	2.158±0.765^△△	276.85±50.67^△△	189.96±30.21^△△
High dose	1.864±0.919^△△	159.72±56.91^△△	109.85±26.91^△△

表儿茶素对对乙酰氨基酚诱导小鼠肝损伤的改善作用及其机制

Improvement effect of epicatechin on liver injury in mice induced by acetaminophen and its mechanism

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