Lianhe Xiaozhi ointment ameliorates metabolic dysfunction-associated steatotic liver disease via peroxisome proliferator-activated receptor alpha pathway activation

BACKGROUND Lianhe Xiaozhi ointment (LXO), an innovative formulation derived from the classic Huanglian Wendan decoction, has been granted a national invention patent. With increasing years of clinical expertise within the Jiangsu Province Hospital of Chinese Medicine in China, LXO has become increasingly recognized as a potent remedy for metabolic disorders, particularly metabolic dysfunction-associated steatotic liver disease (MASLD). To date, specific bioactive components and underlying mechanisms remain unclear. AIM To determine the bioactive components of LXO and clarify its mechanisms of action relevant to management of MASLD. METHODS We used ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry and network pharmacology approaches to systematically determine the key bioactive components of LXO and elucidate biological pathways modulated in the treatment of MASLD. Critical signaling pathways were also elucidated via hepatic transcriptomic analysis, and an exhaustive and rigorous exploration of the therapeutic efficacy and underlying mechanisms of LXO in MASLD was conducted via a combination of in vivo and in vitro high-fat model experiments. RESULTS Network pharmacology analysis revealed six pivotal bioactive components within LXO that collectively serve as the cornerstone for their efficacy against MASLD. LXO exerts multiple therapeutic effects, including weight gain retardation, amelioration of glucose and lipid metabolism disturbances, liver injury mitigation, hepatic inflammation alleviation, and correction of gut microbiota disorders. Multiple platform analyses, including hepatic transcriptomics, quantitative real-time polymerase chain reaction, and western blotting, confirmed that LXO induces peroxisome proliferator-activated receptor alpha (PPARα) transcriptional activation in the liver. In vitro studies confirmed that a PPARα antagonist markedly diminishes the lipid-lowering capacity of LXO. CONCLUSION Collectively, our findings suggest that LXO exerts therapeutic effects on high-fat diet-induced MASLD at least in part via PPARα pathway activation.