Pien-Tze-Huang ameliorates autoimmune hepatitis by promoting hepatocyte autophagy via inhibiting miR-342-3p/mTOR signal.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Pien Tze Huang (PTH), as a national protected species with a history of more than 450 years, is known for its efficacy in heat clearance, detoxification, blood cooling, blood stasis removal, and is commonly used in the treatment of liver-related diseases. However, it remains unclear whether PTH regulates autophagy by modulating miR-342-3p/mTOR signaling pathway for the treatment of autoimmune hepatitis (AIH). AIM OF THE STUDY: To explore the therapeutic effects and molecular mechanisms of PTH on AIH, focusing on the analysis of its regulatory effects on miR-342-3p/mTOR pathway and autophagy. MATERIALS AND METHODS: The chemical composition of PTH was analyzed by LC-MS/MS; AIH model was constructed in C57BL/6J mice to assess the effects of PTH on symptoms, pathological damage and cytokines; lysosomal changes were observed by transmission electron microscopy, and autophagy-related proteins (LC3B, Beclin-1) expression was detected; differential miRNAs and mRNAs were screened by whole-transcriptome sequencing and verified by RT-PCR for key miRNAs and mRNAs (miR-342-3p); induction of AML-12 hepatocyte inflammation model, combined with autophagy modeling experiments, lentivirus-mediated overexpression/knockdown of miR-342-3p and mTOR pathway interventions, to further clarify the molecular mechanism of PTH for AIH. RESULTS: PTH is mainly composed of amino acids and their derivatives, lipids and organic acids; PTH intervention significantly improved the symptoms of AIH mice, attenuated liver injury, regulated cytokines, and promoted lysosomal restitution and autophagy protein expression. Whole transcriptome sequencing and experiments confirmed that PTH downregulated miR-342-3p, inhibited mTOR signaling and enhanced autophagy. Cellular experiments further demonstrated that PTH activated autophagy and alleviated inflammation by directly regulating the miR-342-3p/mTOR axis. CONCLUSION: Pien-Tze-Huang ameliorates autoimmune hepatitis by promoting hepatocyte autophagy via inhibiting miR-342-3p/mTOR signal. This study elucidates a clear molecular mechanism for PTH's therapeutic action, highlighting its potential as a targeted treatment for AIH, and the miR-342-3p/mTOR axis serves as a viable target for future anti-AIH drug design.