Tubuloside B alleviates con A-induced acute liver injury by inhibiting the IL-23/JAK2/STAT3 signaling pathway hyperactivation during Th17 cell differentiation

Tubuloside B (TB), a phenylethanoid glycoside from Cistanche salsa, has been reported to exhibit anti-inflammatory and anti-HCC properties; however, its therapeutic efficacy and underlying mechanisms in concanavalin A (Con A)-induced liver injury remain unclear. We employed a Con A-induced acute liver injury (ALI) mouse model and in vitro Th17 differentiation assays were employed to investigate the therapeutic potential and mechanisms of TB through serum biochemistry, histopathology, flow cytometry, RNA sequencing, qPCR, and Western blotting (WB). TB significantly reduced Con A-induced elevations in serum ALT/AST levels, ameliorated hepatic necrosis and inflammatory infiltration, and suppressed pro-inflammatory cytokines in vivo. RNA-seq and pathway analysis revealed that TB inhibited the IL-17A signaling pathway and restored the Th17/Treg balance by reducing Th17 cell proportions (CD4 + IL-17A + and CD4 + RORγt + ) while increasing Treg proportions (CD25 + Foxp3 + ). Mechanistically, TB inhibited IL-17A-driven activation of NF-κB and MAPK pathways in the liver. Furthermore, molecular docking and in vitro Th17 differentiation assays demonstrated that TB directly targeted IL-23R (ΔG = −10.9 kcal/mol), suppressed IL-23-induced overactivation of the JAK2/STAT3 pathway (p-JAK2, p-STAT3), and inhibited pathogenic Th17 differentiation. Collectively, our findings demonstrate that TB alleviates Con A-induced ALI by blocking the IL-23/JAK2/STAT3 signaling axis during Th17 cell differentiation, thereby attenuating IL-17-mediated hepatic inflammation. This study highlights TB as a potential natural therapeutic candidate for ALI and suggests its potential application in mitigating autoimmune hepatitis (AIH) progression. Mechanism diagram. Exogenous Con A triggers an inflammatory cytokine burst in vivo. These cytokines induce RORγt transcription through JAK2 phosphorylation and subsequent nuclear STAT3 phosphorylation, leading to increased IL-17A expression and secretion. Secreted IL-17A acts on hepatocytes, activating the MAPK/NF-κB signaling pathway and inducing hepatocyte apoptosis. Exogenous TB, however, blocks the IL-23R/JAK2/STAT3 signaling pathway at its source, effectively reducing hepatocyte apoptosis. • Tubuloside B (TB) ameliorates Con A-induced acute liver injury and hepatic inflammation. • TB demonstrates no observable toxicity in healthy mice at therapeutic doses. • Molecular docking identifies TB as a potent IL-23R antagonist (ΔG = −10.9 kcal/mol). • TB mechanistically inhibits IL-23-induced JAK2/STAT3 hyperactivation and pathogenic Th17 cell polarization. • Consequently, TB downregulates the IL-17A-driven NF-κB/MAPK axis, thereby protecting hepatocytes from inflammatory apoptosis.