Mechanistic Study on the Active Components of Bawei Longzuan Granules in Inhibiting Neovascularization

Objective: To explore the active molecular basis and underlying mechanisms of Bawei Longzuan Granules (BWLZ) in inhibiting pannus neovascularization for the treatment of rheumatoid arthritis (RA) through vitro cellular experiments. Methods: Using an inflammation model of HUVECs (human umbilical vein endothelial cells) in vitro, the effects of BWLZ's key active components were validated through CCK-8 assays, flow cytometry, Transwell invasion assays, scratch wound healing assays, and Western blot analyses to elucidate their anti-angiogenic mechanisms in the treatment of RA. Results: CCK-8 and flow cytometry results demonstrated that these compounds significantly inhibited HUVEC proliferation and promoted apoptosis ( P < 0.05). Western blot analysis revealed that VEGF165 markedly upregulated the expression of KDR (VEGFR2) and VEGF in HUVECs ( P < 0.05), whereas naringenin, luteolin, and liquiritigenin reversed VEGF165-induced increases in KDR and VEGFA expression ( P < 0.05). Scratch wound healing assays showed that bevacizumab, liquiritigenin, naringenin, and luteolin significantly reduced VEGF165-induced cell migration rates ( P < 0.05). Transwell invasion assays revealed that VEGF165 significantly enhanced cell invasion compared to the control group ( P < 0.05), while bevacizumab, liquiritigenin, naringenin, and luteolin significantly attenuated VEGF165-induced invasion ( P < 0.05). Tube formation assays demonstrated that VEGF165 significantly promoted the tube formation ability of HUVECs ( P < 0.05), whereas naringenin, luteolin, and liquiritigenin significantly inhibited VEGF165-induced tube formation ( P < 0.05). Conclusion: Three key compounds in BWLZ—luteolin, liquiritigenin, and naringenin—effectively reduced VEGF-induced HUVEC migration, invasion, and tube formation. This anti-angiogenic effect may be associated with the downregulation of VEGFR2 protein expression.