The protective effects of neferine against paracetamol-induced liver injury are associated with the activation of SIRT1/Nrf2/HO-1 signaling pathway and inhibition of NF-kappa B/TNF-alpha/iNOS/COX-II cascade

Drug-induced hepatotoxicity is a significant public health issue that influences the development of novel pharmaceutical therapies and the retraction of numerous promising medications from the market. Therefore, the current study investigated the potential hepato-protective benefits of NEF against hepatotoxicity caused by paracetamol (APAP) in mice and assessed its underlying mechanisms. Mice were divided randomly into six groups; control (received normal saline), NEF control, APAP, N-acetylcysteine (NAC; served as a standard treatment) + APAP, NEF (10 mg/kg) + APAP, and NEF (20 mg/kg) + APAP. The serum and hepatic tissues were collected for different biochemical, genetic, and histological assessments. APAP induced profound hepatic damage that was evident through all biochemical, histological, and molecular assessments. NEF pretreatment opposed the elevation of liver injury biomarkers and attenuated hepatic histological disruption. At the molecular level, NEF increased the hepatic level and protein expression of SIRT-1. NEF increased the hepatic mRNA and protein expression of Nrf2 and HO-1. NEF also decreased hepatic level of oxidative stress biomarker, MDA and increased the hepatic levels of antioxidants: GSH, GR, GST, TAC, and SOD, NEF also counteracted the activation of NF-κB and inhibited the upregulation of different inflammatory cytokines as TNF-α and interleukins- (IL-1β and IL-6). Furthermore, NEF pretreatment decreased the hepatic level and mRNA expression of COX-II and iNOS. NEF ameliorated APAP-induced liver injury in mice where the higher dose of NEF (20 mg/kg) was more effective than the lower (10 mg/kg) compared to NAC. This effect is association with upregulation of SIRT-1/Nrf2/HO-1 and interruption of NF-κB/cytokines/iNOS/COX-II signaling cascades. NEF ameliorated APAP-induced liver injury in mice.NEF increased SIRT-1 hepatic levels and protein expression.NEF increased the mRNA and protein expression of Nrf2 and HO-1.NEF decreased MDA and increased the antioxidants GSH, GR, GST, TAC, and SOD.NEF inhibited the upregulation of NF-κB, TNF-α, IL-1β, and IL-6.NEF pretreatment decreased mRNA expression of COX-II and iNOS. NEF ameliorated APAP-induced liver injury in mice. NEF increased SIRT-1 hepatic levels and protein expression. NEF increased the mRNA and protein expression of Nrf2 and HO-1. NEF decreased MDA and increased the antioxidants GSH, GR, GST, TAC, and SOD. NEF inhibited the upregulation of NF-κB, TNF-α, IL-1β, and IL-6. NEF pretreatment decreased mRNA expression of COX-II and iNOS.

药物性肝损伤（Drug-induced hepatotoxicity）是一类重大公共卫生问题，可阻碍新型药物治疗方案的研发，并导致多款极具潜力的药物从市场撤市。为此，本研究探究了NEF对乙酰氨基酚（paracetamol, APAP）诱导的小鼠肝损伤的潜在保肝作用，并对其作用机制进行了系统分析。实验将小鼠随机分为6组：正常对照组（给予生理盐水）、NEF对照组、APAP模型组、N-乙酰半胱氨酸（N-acetylcysteine, NAC，作为阳性对照药）+APAP组、低剂量NEF（10 mg/kg）+APAP组以及高剂量NEF（20 mg/kg）+APAP组。采集血清与肝组织样本，开展生化、遗传及组织病理学等多维度检测。APAP可诱导严重的肝损伤，该损伤在生化、组织学及分子层面的检测中均得到验证。NEF预处理可逆转肝损伤生物标志物的升高，并减轻肝组织的病理学破坏。在分子层面，NEF可提升肝组织中沉默信息调节因子1（SIRT-1）的水平与蛋白表达量；上调核因子E2相关因子2（Nrf2）与血红素氧合酶1（HO-1）的mRNA及蛋白表达水平；降低肝组织中氧化应激标志物丙二醛（MDA）的含量，同时提升谷胱甘肽（GSH）、谷胱甘肽还原酶（GR）、谷胱甘肽S-转移酶（GST）、总抗氧化能力（TAC）与超氧化物歧化酶（SOD）等抗氧化指标的水平。此外，NEF可抑制核因子κB（NF-κB）的活化，并下调肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）与白细胞介素6（IL-6）等多种炎症细胞因子的表达；同时降低环氧合酶2（COX-II）与诱导型一氧化氮合酶（iNOS）的肝组织水平及mRNA表达量。与NAC相比，高剂量NEF（20 mg/kg）的保肝效果优于低剂量NEF（10 mg/kg）。本研究证实，NEF可改善APAP诱导的小鼠肝损伤，其作用机制与激活SIRT-1/Nrf2/HO-1通路、阻断NF-κB/细胞因子/iNOS/COX-II信号级联反应密切相关。