Regulatory effect of Pien Tze Huang on the mTreg/mTh17 balance in mice with autoimmune hepatitis

• PTH attenuates ConA-induced autoimmune hepatitis in mice. • PTH regulates the balance between memory Treg/Th17 cells. • PTH effectively inhibits glycolytic pathway activation To investigate the therapeutic efficacy and underlying mechanisms of Pien Tze Huang (PTH) in a murine autoimmune hepatitis (AIH) model through assessment of memory regulatory T cells (mTreg)/memory T helper cell 17 (mTh17) cell equilibrium and glycolytic metabolism. AIH was induced through intravenous concanavalin A (ConA) administration via tail vein injection with prophylactic PTH treatment over 10 days. Hepatic histopathological alterations were evaluated using H&E staining, while serum transaminase levels were quantified through biochemical analysis. ELISA was employed to determine immunoglobulin concentrations and hepatic tissue levels of interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 21 (IL-21), interleukin 10 (IL-10), interleukin 4 (IL-4), tumor necrosis factor α (TNF-α), interferon-gamma (IFN-γ), Immunoglobulin G (IgG), Immunoglobulin A (IgA), Immunoglobulin M (IgM), glycolysis hexokinase 2 (HK2), pyruvate kinase (PK), glucose-6-phosphatase (G6pase), phosphoenolpyruvate carboxykinase (PEPCK), aldolase A (ALDOA), glucokinase (GCK), lactate dehydrogenase A (LDHA). Flow cytometric analysis was performed to quantify Treg, mTreg, Th17, and mTh17 cell populations. Protein expression of glucose transporter protein 1 (Glut1), glucose transporter protein 2 (Glut2), glucose transporter protein 3 (Glut3), glucose transporter protein 4 (Glut4), hypoxia-inducible factor-1α subunit (HIF-1α), signal transducer and activator of transcription 3 (STAT3), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), T-cell transcription factor (T-bet), and retinoic acid receptor-related orphan receptor gamma (RORγt) was analyzed by Western blotting. Relative to the model group, PTH administration significantly ameliorated hepatocellular injury, as evidenced by reduced serum ALT and AST levels, decreased immunoglobulin concentrations, and attenuated hepatic pathological damage ( p < 0.05 or p < 0.01). Furthermore, PTH treatment resulted in significant suppression of proinflammatory cytokine expression in murine hepatic tissue ( p < 0.05 or p < 0.01). PTH treatment increased the proportions of Treg and mTreg cells while reducing Th17 and mTh17 cell populations ( p < 0.05 or p < 0.01). Mechanistic investigations revealed that PTH significantly downregulated the expression of critical glycolytic enzymes and glycolysis-associated proteins in hepatic tissue of AIH model mice ( p < 0.05 or p < 0.01). PTH effectively attenuates ConA-induced AIH by modulating the balance of memory Treg/Th17 cells and glycolytic pathways.