Shen Xing-Ling, Guo Yan-Na, Lu Meng-Han, Ding Kang-Ning, Liang Shao-Shan, Mou Rui-Wei, Yuan Sheng, He Yong-Ming, Tang Lu-Ping
School of Life Science and Engineering, Foshan University, Foshan 528225, China.
School of Life Science and Engineering, Foshan University, Foshan 528225, China.
Ecotoxicol Environ Saf. 2023 Mar 1;252:114590. doi: 10.1016/j.ecoenv.2023.114590. Epub 2023 Feb 2.
To explore the action time and molecular mechanism underlying the effect of acetaminophen (APAP) on liver injury. APAP was used to establish drug-induced liver injury (DILI) model in mice. Mice in the model group were intraperitoneally injected 300 mg/kg APAP for 6, 12, and 24 h respectively, and control group mice were given the same volume of normal saline. The mice were anesthetized through intravenous injection of sodium pentobarbital at 6, 12, and 24 h after APAP poisoning. Analysis of ALT, AST and ALP in serum, liver histopathological observation, oxidative damage and western blot were performed. The livers in APAP exposed mice were pale, smaller, with a rough texture, and poorly arranged cells. Lesions, large areas of hyperemia, inflammation, swelling, poorly cell arrangement, necrosis, and apoptosis of liver cells were obvious in the liver tissue sections. Serum ALT, AST and ALP levels were significantly enhanced at 12 h of APAP adminstration mice than that of in control group mice (P＜0.05). The histopathological alterations and proinflammatory cytokines (IL-1β, TNF-α and IL-6) levels were most severe at 12 h of APAP-induced hepatotoxicity. APAP treatment induced oxidative stress by decreasing hepatic activities of superoxide dismutase (SOD) and glutathione (GSH) (P＜0.05), and enhancing malondialdehyde (MDA) content (P＜0.05). Moreover, APAP inhibited erythroid 2-related factor 2 (Nrf2) antioxidative pathway with decreased of Nrf2 and HO-1 proteins levels. Furthermore, APAP aggravated the activation of NLRP3 inflammasome by increasing of NLRP3, caspase-1, ASC, IL-1β and IL-18 proteins levels. Finally, APAP further significantly activated the toll-like receptor 4 (TLR4), nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) signaling pathways. This study demonstrated that APAP-induced hepatotoxicity by inhibiting of Nrf2 antioxidative pathway and promoting TLR4-NF-κB-MAPK inflammatory response and NLRP3 inflammasome activation.
探讨对乙酰氨基酚（APAP）致肝损伤作用的时间及分子机制。采用APAP建立小鼠药物性肝损伤（DILI）模型。模型组小鼠分别腹腔注射300mg/kg APAP，持续6、12和24小时，对照组小鼠给予相同体积的生理盐水。在APAP中毒后6、12和24小时，通过静脉注射戊巴比妥钠对小鼠进行麻醉。检测血清谷丙转氨酶（ALT）、谷草转氨酶（AST）和碱性磷酸酶（ALP），进行肝脏组织病理学观察、氧化损伤分析及蛋白质免疫印迹法检测。APAP处理后的小鼠肝脏颜色变浅、体积变小、质地粗糙、细胞排列紊乱。肝脏组织切片可见病变，大面积充血、炎症、肿胀、细胞排列紊乱、坏死及肝细胞凋亡。APAP给药12小时的小鼠血清ALT、AST和ALP水平显著高于对照组小鼠（P＜0.05）。APAP诱导肝毒性12小时时，组织病理学改变及促炎细胞因子（IL-1β、TNF-α和IL-6）水平最为严重。APAP处理通过降低肝脏超氧化物歧化酶（SOD）和谷胱甘肽（GSH）活性诱导氧化应激（P＜0.05），并增加丙二醛（MDA）含量（P＜0.05）。此外，APAP抑制红系2相关因子2（Nrf2）抗氧化途径，使Nrf2和血红素加氧酶-1（HO-1）蛋白水平降低。此外，APAP通过增加NLRP3、半胱天冬酶-1（caspase-1）、凋亡相关斑点样蛋白（ASC）、IL-1β和IL-18蛋白水平加重NLRP3炎性小体的激活。最后，APAP进一步显著激活Toll样受体4（TLR4）、核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPKs）信号通路。本研究表明，APAP通过抑制Nrf2抗氧化途径及促进TLR4-NF-κB-MAPK炎症反应和NLRP3炎性小体激活诱导肝毒性。
