Berberine Enhances the Immunomodulatory Function of Canine Bone Marrow Mesenchymal Stem Cells via Regulation of the PD-1/PD-L1 Pathway: A Network-Pharmacology Analysis and In Vitro Experimental Validation

As a result of network pharmacology screening, 82 common targets of berberine were identified in dog BMSCs. KEGG pathway enrichment analysis showed that 9 key targets of berberine, including PD-L1, PD-L2, STAT3, AKT1, JAK2, MTOR, TLR4, RELA, and MAPK14, were enriched in the PD-1/PD-L1 immune pathway; CCK-8 and SA - β - gal analysis showed that treatment with 0-20 μ M berberine for 48 hours did not significantly reduce the proliferation activity of BMSCs in dogs; The ELISA results showed that berberine at a concentration of 10-20 μ M for 48 hours promoted the synthesis and secretion of sPD-L1 and sPD-L2, while inhibiting the synthesis and secretion of IL-6 and TNF - α, with no significant effect on IL-4 and IL-10.

The RT qPCR results showed that after treatment with berberine, the expression level of PD-L1 gene was upregulated, TLR4, RELA, and MAPK14 were downregulated, while the expression levels of STAT3, AKT1, JAK2, and MTOR remained unchanged. Conclusion: Berberine enhances the immune regulatory function of canine BMSCs through the PD-1/PD-L1 pathway, providing a theoretical basis for the clinical application of canine BMSCs and their combination with berberine.