Alsahli Tariq G, Khalid Maryam, Malik Muhammad Nasir Hayat, Alshammari Saud O
Department of Pharmacology, College of Pharmacy, Jouf University, Sakaka 72341, Saudi Arabia.
Faculty of Pharmacy, The University of Lahore, Lahore 54000, Pakistan.
Medicina (Kaunas). 2025 Jun 6;61(6):1048. doi: 10.3390/medicina61061048.
Alcoholic liver disease (ALD) is a major health burden caused by chronic alcohol consumption, leading to oxidative stress, inflammation, and fibrosis. Current treatments are limited, highlighting the need for novel therapeutic agents. This study investigated the hepatoprotective effects of 'Terpinen-4-ol (T4OL)', a natural monoterpene from tea tree oil, against ethanol-induced liver injury, focusing on its molecular and cellular mechanisms. Network pharmacology and molecular docking were employed to predict T4OL's interaction with ALD-associated targets. Human HepG2 cells were used to validate the in silico findings. Cells were exposed to ethanol (8%) prior to treatment with T4OL or silymarin (SIL), and cytotoxicity was assessed through MTT, crystal violet, and trypan blue assays. Moreover, ELISA and qPCR were conducted to evaluate antioxidant, inflammatory, and fibrotic markers. Network pharmacology analysis suggested that T4OL exerts its hepatoprotective effects by suppressing inflammatory and fibrotic mediators (HIF-1α, TGF-β1, and TNF-α). Docking studies also exhibited a strong binding affinity of T4OL to key ALD targets, with docking scores comparable to SIL. In addition, T4OL (13-1300 µM) dose-dependently protected HepG2 cells from ethanol-induced damage, restoring viability by up to 80% at 650 µM. It significantly elevated antioxidant levels (GSH by 2.5-fold, SOD by 1.8-fold) and suppressed pro-inflammatory and fibrotic markers (IL-6, COL1A1, TIMP-1) by 40-60%. At higher concentrations (650-1300 µM), T4OL outperformed SIL in cytoprotection and anti-fibrotic effects. T4OL mitigates ethanol-induced liver injury by targeting oxidative stress, inflammation, and fibrosis pathways, demonstrating superior efficacy to SIL at optimal doses. Its multi-target action supports its potential as a therapeutic candidate for ALD.
酒精性肝病（ALD）是由长期饮酒导致的主要健康负担，会引发氧化应激、炎症和纤维化。目前的治疗方法有限，这凸显了新型治疗药物的必要性。本研究调查了茶树油中的天然单萜“萜品烯 - 4 - 醇（T4OL）”对乙醇诱导的肝损伤的保肝作用，重点关注其分子和细胞机制。采用网络药理学和分子对接来预测T4OL与ALD相关靶点的相互作用。使用人肝癌细胞系HepG2细胞来验证计算机模拟研究结果。在用T4OL或水飞蓟宾（SIL）处理之前，将细胞暴露于乙醇（8%）中，并通过MTT、结晶紫和台盼蓝试验评估细胞毒性。此外，进行酶联免疫吸附测定（ELISA）和定量聚合酶链反应（qPCR）以评估抗氧化、炎症和纤维化标志物。网络药理学分析表明，T4OL通过抑制炎症和纤维化介质（缺氧诱导因子 - 1α、转化生长因子 - β1和肿瘤坏死因子 - α）发挥其保肝作用。对接研究还显示T4OL与关键的ALD靶点具有很强的结合亲和力，对接分数与SIL相当。此外，T4OL（13 - 1300 μM）呈剂量依赖性地保护HepG2细胞免受乙醇诱导的损伤，在650 μM时可将细胞活力恢复高达80%。它显著提高抗氧化水平（谷胱甘肽增加2.5倍，超氧化物歧化酶增加1.8倍），并将促炎和纤维化标志物（白细胞介素 - 6、I型胶原蛋白α1链、金属蛋白酶组织抑制因子 - 1）抑制40 - 60%。在较高浓度（650 - 1300 μM）下，T4OL在细胞保护和抗纤维化作用方面优于SIL。T4OL通过靶向氧化应激、炎症和纤维化途径减轻乙醇诱导的肝损伤，在最佳剂量下显示出优于SIL的疗效。其多靶点作用支持其作为ALD治疗候选药物的潜力。
