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

Background

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.

Methods

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).

Results

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.

Conclusions

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.