Astragaloside IV Pretreatment Boosts BMSC Efficacy in Murine Fulminant Hepatic Failure by Enhancing Stemness and Reducing Apoptosis

Fulminant hepatic failure (FHF) is a severe condition characterized by rapid liver cell death and high mortality, with limited effective treatments beyond liver transplantation. Bone marrow-derived mesenchymal stem cells (BMSCs) offer promising regenerative potential for liver repair due to their immunomodulatory and differentiation capabilities. However, the harsh, inflammatory, and hypoxic microenvironment of the injured liver significantly compromises BMSC viability and therapeutic efficacy upon transplantation. There is a critical need for strategies to enhance BMSC resilience and regenerative capacity in such adverse conditions, making preconditioning approaches a key area of research.

Researchers established a murine model of fulminant hepatic failure (FHF) using intraperitoneal injection of D-galactosamine and lipopolysaccharide (D-GalN/LPS). Bone marrow mesenchymal stem cells (BMSCs) were cultured and pretreated with Huangqi injection (HQI) to create HQI-BMSCs, or with Astragaloside IV (AS-IV), a key compound in HQI. Mice were divided into four groups: normal control, FHF, FHF + untreated BMSCs, and FHF + HQI-BMSCs. BMSCs were immediately transplanted via tail vein post-FHF induction. At 4 hours post-transplantation, serum levels of ALT, TBIL, IL-10, and IL-6 were measured. Liver tissue underwent hematoxylin-eosin staining for histopathology, TUNEL assay for apoptosis, and immunohistochemistry for PCNA (proliferating cell nuclear antigen). PCNA protein expression was also quantified by western blotting. CCK-8 assay assessed BMSC viability, and qPCR was used for stemness-related gene expression.

Pretreatment with Huangqi injection (HQI) markedly improved BMSC viability, suggesting enhanced cellular resilience. In the D-GalN/LPS-induced FHF model, transplantation of HQI-pretreated BMSCs significantly mitigated liver injury. Specifically, HQI-BMSCs reduced serum levels of ALT, TBIL, and the pro-inflammatory cytokine IL-6, while also suppressing hepatocyte apoptosis in liver tissue. Furthermore, HQI-BMSCs conferred greater hepatoprotective effects compared to untreated BMSCs, leading to improved biochemical indices and reduced histopathological damage. Investigation into the active constituent, Astragaloside IV (AS-IV), revealed its ability to enhance the expression of stemness-related genes in BMSCs. AS-IV also promoted G1-to-S/G2 phase progression in BMSCs, indicating improved cell cycle activity and proliferative capacity. Importantly, AS-IV attenuated LPS-induced apoptosis in BMSCs, directly addressing a key challenge of BMSC survival in inflammatory environments. > HQI-BMSCs demonstrated superior therapeutic efficacy in reducing liver injury markers and histopathological damage compared to untreated BMSCs in the FHF model.