Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) <i>via</i> dephosphorylation of EGFR signaling pathway

This study aims to assess the impact of Timosaponin AⅢ (T-AⅢ) on drug-metabolizing enzymes in anticancer treatment. In vivo experiments were conducted in nude mice and ICR mice. Following 24 days of T-AⅢ administration, nude mice exhibited induction of CYP2B10, MDR1, and CYP3A11 in the liver. In the liver of ICR mice, CYP2B10 and MDR1 were up-regulated after 3 days of T-AⅢ administration. In vitro assessments were conducted using HepG2 cells to ascertain the effects and underlying mechanisms. In HepG2 cells, T-AⅢ induced the expression of CYP2B6, MDR1, and CYP3A4, along with CAR activation. CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4. Furthermore, other CAR target genes displayed significant up-regulation. Up-regulation of mCAR was observed in the livers of nude mice and ICR mice. Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation, partially reversed by the MAPK/MEK activator t-BHQ. Inhibition of the ERK1/2 signaling pathway was also observed in vivo. Lastly, T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845, and it suppressed EGF-induced phosphorylation of EGFR, ERK, and CAR. Additionally, T-AⅢ inhibited EGFR phosphorylation in nude mice. Our results demonstrated that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.