IRAK4在对乙酰氨基酚诱导的急性肝细胞损伤中的作用及其机制

doi:10.3969/j.issn.1004-616x.2022.04.002

摘要/Abstract

摘要： 目的：探讨白细胞介素-1受体相关激酶4(IRAK4)在对乙酰氨基酚(APAP)诱导急性肝L02细胞损伤过程中的作用及其机制。方法：将人正常肝细胞系L02分为对照组，20 mmol/L APAP处理6、12和24 h组，分别采用CCK-8法检测细胞活力，AnnexinV/PI双染法检测细胞凋亡率，MitoSOX和JC-1荧光染料检测线粒体活性氧水平和膜电位变化，荧光定量PCR (qPCR)和Western blot检测IRAK4的相对表达量。在沉默IRAK4基因后按前述方法检测L02细胞活力、凋亡和线粒体活性氧变化。进一步给予10 mmol/L N-乙酰半氨酸(NAC)干预24 h后，用CCK-8法检测L02细胞活力，qPCR检测IRAK4 mRNA表达水平。结果：与对照组相比，20 mmol/L APAP处理L02细胞24 h后，细胞活力下降且可明显诱导细胞发生凋亡(P<0.05)；20 mmol/L APAP处理12和24 h后线粒体活性氧水平明显升高(P<0.05)；不同时间点线粒体膜电位均显著降低(P<0.05)；qPCR和Western blot检测结果显示，20 mmol/L APAP处理24 h后IRAK4表达水平升高(P<0.05)。与对照组和APAP组相比，沉默IRAK4基因后，细胞活力增加、凋亡程度减轻以及线粒体活性氧降低(P<0.05)。与APAP组相比，给予NAC后细胞活力增加，IRAK4 mRNA表达水平降低(P<0.05)。结论：IRAK4可能参与了APAP诱导的急性肝细胞损伤，是APAP致肝细胞损伤的一个潜在治疗靶点。

Abstract: OBJECTIVE: To investigate the role and mechanism of IRAK4 in acute liver cell injury induced by acetaminophen (APAP). METHODS: Human liver cell line L02 cells were divided into control and treatment groups. The latter were treated with 20 mmol/L APAP for 6,12 and 24 hours. Cell viability was detected by CCK-8 method,apoptosis rates were detected by flow cytometry combined with AnnexinV/PI double staining, mitochondrial changes were detected by MitoSOX and JC-1 fluorescence dye, relative expression levels of IRAK4 were detected by fluorescence quantitative PCR (qPCR) and Western blot,L02 cell viability, and apoptosis and mitochondrial reactive oxygen species (mtROS) were detected after silencing the IRAK4 gene. After treatment with 10 mmol/L N-acetylhemionine (NAC) for 24 hours,L02 cell viability was detected by CCK-8 method,and IRAK4 gene expression level was detected by qPCR. RESULTS: Compared with the control group,cell viabilities were reduced and apoptosis rates were significantly increased after the APAP treatment for 24 h (P<0.05). After 12 and 24 h treatments,the levels of mtROS increased significantly (P<0.05);mitochondrial membrane potential decreased significantly at different time points (P<0.05). qPCR and Western blot results showed that IRAK4 expression increased after treatment for 24 h (P<0.05). After silencing IRAK4 gene,compared with the control and APAP groups,cell viability increased,apoptosis decreased and mtROS decreased (P<0.05). Compared with APAP group,NAC treatment increased cell viability and decreased mRNA expression of IRAK4 gene (P<0.05). CONCLUSION: IRAK4 might be involved in APAP-induced acute liver cell injury and would be a therapeutic target of APAP liver cell injury.

Key words: IRAK4, acetaminophen, oxidative stress, liver injury

中图分类号:

R394.6

蔡娇, 孔德钦, 龙子, 彭洁, 刘江正, 刘瑞, 海春旭. IRAK4在对乙酰氨基酚诱导的急性肝细胞损伤中的作用及其机制[J]. 癌变·畸变·突变, 2022, 34(4): 255-261.

CAI Jiao, KONG Deqin, LONG Zi, PENG Jie, LIU Jiangzheng, LIU Rui, HAI Chunxu. Role of IRAK4 in acetaminophen-induced acute liver cell injury[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(4): 255-261.

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