Total astragalus saponins enhance the anti-liver fibrosis efficacy of Numb-overexpressed hUC-MSCs through stabilizing the Numb-p53 axis.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragali Radix (Huangqi), a traditional Chinese medicine documented in Shennong Bencao Jing (ca. 200 AD), has been used for chronic liver disease. Total astragalus saponins (TAS), its primary bioactive component, exhibit anti-fibrotic effects. However, their potential synergy with stem cell therapy remains unexplored.

AIM OF THE STUDY: To investigate whether TAS enhances the therapeutic efficacy of Numb-overexpressing human umbilical cord-mesenchymal stem cells (hUC-MSCs Numb-OE) against cholestatic liver fibrosis (CLF) and elucidate underlying mechanisms.

MATERIALS AND

METHODS

CLF was induced in rats by bile duct ligation, post-surgery, rats received hUC-MSCs Numb-OE via intrasplenic injection. Starting the second week, TAS was administered daily by gavage for three weeks. Serum biochemistry, liver histopathology, hepatic hydroxyproline content, ductular reaction (DR), liver regeneration and Numb-p53 axis were assessed. In vitro, the effect of TAS on hUC-MSCs Numb-OE differentiation was evaluated.

RESULTS

Combined TAS and hUC-MSCs Numb-OE significantly improved inflammatory response and cholestasis, reduced collagen deposition, and inhibited hepatic stellate cells activation and DR. Furthermore, the combination promoted differentiation of hUC-MSCs Numb-OE into hepatocytes confirmed by enhanced expression of hepatocyte nuclear factor 4α and albumin, and by EGFP (marked hUC-MSCs)/albumin immunofluorescent co-staining in vitro. Mechanistically, the combined therapy enhanced stability of the Numb-p53 axis demonstrated by increased expression levels of Numb and p53, alongside reduced expression levels of pNumb and Mdm2.

CONCLUSIONS

TAS synergistically enhances the anti-CLF efficacy of hUC-MSCs Numb-OE. This effect is associated with enhanced suppression of DR and promotion of liver regeneration, may mediated through stabilization of the Numb-p53 axis.