Aqueous Extract of Lonicerae japonicae Flos Attenuates Alcohol-Associated Liver Disease by Suppressing Oxidative Stress and Inflammation in Mice

Abstract

Objectives: This study employed an integrative strategy combining UPLC-MS-based phytochemical profiling, network pharmacology, and transcriptomics to systematically investigate the protective effects and potential mechanisms of LJF against ALD. Methods: An ALD model was established via chronic ethanol liquid diet feeding combined with acute ethanol binge administration over 16 days. Liver function, lipid metabolism, oxidative stress (OS), and inflammatory markers were assessed. UHPLC–Q Exactive HFX MS analysis was performed to tentatively annotate the chemical constituents of LJF. Candidate targets and active constituents were predicted by network pharmacology integrated with transcriptomics, followed by experimental validation using quantitative real-time PCR (qRT-PCR) and Western blot. Results: LJF extract significantly alleviated ALD, as evidenced by decreased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, reduced hepatic total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) levels, and improved histopathological changes. LJF attenuated oxidative stress by lowering reactive oxygen species (ROS) and malondialdehyde (MDA) levels while elevating superoxide dismutase (SOD) and glutathione (GSH) levels. Furthermore, LJF suppressed inflammatory responses by reducing IL-6, IL-1β, and TNF-α levels. Integrative analyses pinpointed Hif1a, Ppara, Cyp2b10, Chac1, Nupr1 and Saa1/2, among others, as key targets, with luteolin, apigenin, isorhamnetin, lonicerin, 5-O-caffeoyl shikimic acid, and caffeic acid as major bioactive constituents. Mechanistically, LJF concurrently restored the Nrf2/HO-1 antioxidant defense and suppressed the p38 MAPK/NF-κB pro-inflammatory axis, thereby exerting a dual regulatory effect on redox homeostasis and immune response. Conclusions: These findings establish LJF as a promising multi-target agent against ALD and, more importantly, provide correlative evidence for the involvement of Nrf2/HO-1 regulation and p38 MAPK/NF-κB signaling modulation, which may guide future strategies for ALD pharmacotherapy.