Danggui-Shaoyao-San ameliorates metabolic dysfunction-associated steatohepatitis via suppressing hepatic macrophage NLRP3 inflammasome.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui-Shaoyao-San (DGSY) is a classic prescription in traditional Chinese medicine that has demonstrated therapeutic efficacy in treating fatty liver. However, its pharmacological mechanisms remain inadequately explored.

AIM OF THE STUDY: This study aims to elucidate the molecular mechanisms through which DGSY may contribute to the modulation of MASH progression.

MATERIALS AND

METHODS

A MASH model was established in mice using a high-fat high-carbohydrate (HFHC) diet, followed by treatment with DGSY to assess its efficacy. RNA sequencing (RNA-Seq) analysis, western blotting, and immunofluorescence were employed to elucidate the potential mechanisms of DGSY in the treatment of MASH. The MASH mouse model and bone marrow-derived macrophages (BMDMs) stimulated with LPS + ATP were utilized to investigate the activation of the NLRP3 inflammasome in macrophages. Additionally, the mechanisms of DGSY were further validated through NLRP3 knockdown in vivo and knockout in vitro. UHPLC-Q-Orbitrap HRMS analysis was employed to identify the primary active components of DGSY.

RESULTS

DGSY was effective in treating the HFHC diet-induced MASH mouse model, significantly alleviating liver inflammation. Mechanistically, RNA-Seq analysis indicated that the NOD-like receptor signaling pathway was significantly affected in DGSY-mediated effects. The protein expression levels of NLRP3, caspase1, and IL-1β were significantly elevated in liver macrophages from the MASH mouse model and in LPS + ATP-stimulated BMDMs, which were reversed by DGSY treatment. NLRP3 knockdown in vivo alleviated MASH but counteracted the therapeutic effects of DGSY. Both NLRP3 knockdown in vivo and knockout in vitro abolished its effect on inhibiting caspase1 activity and IL-1β release. In addition, UHPLC-Q-Orbitrap HRMS analysis identified four main active ingredients in DGSY, with atractenolide III and gallic acid notably inhibiting NLRP3 protein expression.

CONCLUSIONS

DGSY exerts a protective effect against MASH, inhibiting hepatic macrophage NLRP3 inflammasome, suppressing caspase1 activity, and reducing IL-1β release. This study provides experimental evidence for the clinical application of DGSY in the treatment of MASH.