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

吉林大学学报(医学版)

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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:2025-01-26 发布日期:2025-01-26
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:
吉林省教育厅科学研究重点项目(JJKH20241337KJ)

Ameliorating effect of epicatechin on liver injury induced by acetaminophen in mice 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，College 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，Songyuan 138000，China
^6.Department of Radiotherapy，Tumor Hospital，Jilin Province，Changchun 130012，China

Received:2024-12-09 Accepted:2025-01-16 Online:2025-01-26 Published:2025-01-26
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）诱导的小鼠肝损伤的保护作用，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EC组（10 mg·kg^-1）、中剂量EC组（20 mg·kg^-1）和高剂量EC组（40 mg·kg^-1），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。24 h后处死小鼠收集血液和肝脏组织用于后续检测。采用HE染色检测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）水平以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组小鼠血清中ALT及AST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MPO活性及TNF-α和IL-1β水平降低（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<0.01），ATP和GSH水平降低（P<0.01）；与APAP模型组比较，低、中和高剂量小鼠肝组织MDA和Fe²⁺水平明显降低（P<0.05或P<0.01），ATP和GSH水平降低（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<0.01）；与APAP模型组比较，低、中和高剂量小鼠肝组织xCT和GPX4蛋白表达水平明显升高（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<0.01）；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Nrf2和血清素加氧酶1（HO-1）水平升高（P<0.01）；与APAP模型组比较，低、中和高剂量EC组小鼠肝组织中Nrf2和HO-1水平明显升高（P<0.01）。与对照组比较，APAP模型组Nrf2^-/-小鼠血清中ALT水平及肝组织中MDA和Fe²⁺水平明显升高（P<0.01），小鼠肝组织中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 explore the protective effect of epicatechin （EC） on acetaminophen （APAP）- induced liver injury in the mice， and to clarify its possible mechanisms. Methods Sixty C57BL/6J mice were randomly divided into blank control group， APAP model group， low dose of EC group （10 mg·kg^-1）， medium 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 the mice in blank control group， the mice in other groups were intraperitoneally injected with APAP （200 mg·kg^-1） to induce the liver injury models. One hour before APAP injection， the mice in low， medium， and high doses of EC groups were intraperitoneally injected with 10， 20， and 40 mg·kg^-1 EC， respectively.Thirty-six nuclear factor erythroid 2-related factor 2 deficitent mice （Nrf2^-/- mice） were randomly divided into control group， APAP group and APAP+EC group. After 24 h， the mice were sacrificed， and blood and liver tissue were collected for subsequent analysis. The pathomorphology of live tissue of the mice in various groups were assessed by HE staining， while the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， the activities of myeloperoxidase （MPO） in liver tissue， and the levels of tumor necrosis factor-α（TNF-α）， interleukin-1β （IL-1β）， malondialdehyde （MDA）， adenosine triphosphate （ATP）， and glutathione （GSH），and ferrous ions （Fe²⁺） in liver tissve of the mice were determined with kits. The expression levels of nuclear factor-kappa B （NF-κB） and Nrf2 signaling pathway-related proteins in liver tissue of the mice in various groups were detected by Western blotting method. Results Compared with APAP model group， the liver injuries induced by APAP in different doses of EC groups were improved. Compared with blank control group， the serum ALT and AST levels of the mice in APAP model group were significantly increased （P<0.01）； compared with APAP model group， the serum ALT and AST levels in low， medium 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 TNF-α and IL-1β levels in liver tissue of the mice in APAP model group were significantly increased （P<0.01）； compared with APAP model group， the MPO activities and the levels of TNF-α and IL-1β in liver tissue in low， medium 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<0.01）， and the levels of ATP and GSH in liver tissue were decreased （P<0.01）； compared with APAP model group， the levels of MDA and Fe²⁺ in liver tissue of the mice in different doses of EC groups were significantly decreased （P<0.05 or P<0.01）. Compared with blank control group， the GPX4 and xCT protein expression levels in liver tissue of the mice in APAP model group were significantly decreased （P<0.01）； compared with APAP model group， the expression levels of GPX4 and xCT in liver tissue of the mice in different doses of EC were significantly increased（P<0.05 or P<0.01）. Compared with blank control group， the expression levels of NF-κB p-p65 and phosphorylated inhibitor of NF-κB alpha（p-IκBα） in liver tissue of the mice in APAP mode group were significantly increased expression（P<0.01）. Companed with blank control group， the levels of NF-κB p-p65 and p-IκBα proteins in liver tissue of the mice in low， medium and high doses of EC groups were significantly reduced（P<0.01）. Compared with blank control group， the levels of Nrf2 and HO-1 in liver tissue of the mice in APAP model group were increased （P<0.01）； compared with APAP model group， the levels of Nrf2 and HO-1 in liver tissue of the mice in low， medium and high doses of EC groups were significantly increased（P<0.01）. Compared with control group， the serum AST and ATL and level 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 of the mice were significantly decreased（P<0.01）. Conclusion EC can alleviate APAP-induced liver injury， and the mechanism may be related to the activation of Nrf2/GPX4 signaling pathway by EC to inhibit iron death.

Key words: Epicatechin, Acetaminophen, Liver injury, Ferroptosis, nuclear factor erythroid 2-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, (): 1-10.

Huiyuan YU, Ling JIN, Ying YU, Xue WANG, Bing WANG. Ameliorating effect of epicatechin on liver injury induced by acetaminophen in mice and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2025, (): 1-10.

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Group	Serum ALT level [λ_B/（U·L^-1）]	Serum AST level [λ_B/（U·L^-1）]
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^△
Medium 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^△
Medium 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^△△

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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

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

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