Taohong Siwu-Containing Serum Enhances Angiogenesis in Rat Aortic Endothelial Cells by Regulating the VHL/HIF-1α/VEGF Signaling Pathway.

BACKGROUND: The incidence of bone fracture and bone-related diseases is increasing every year. Angiogenesis plays a vital role in fracture healing and bone repair. This study assessed the benefits of Taohong Siwu (TSW) decoction on angiogenesis in isolated rat aortic endothelial cells (RAEC) treated with TSW-containing serum. METHODS: The components of TSW decoction were analyzed by liquid chromatography-mass spectrometry (LC-MS). TSW-containing serum was prepared by gavage of TSW decoction to Sprague-Dawley (SD) rats. The effects of TSW-containing serum on the viability, migration, wound healing, and angiogenesis of RAEC were detected by the MTT, transwell, wound healing, and Matrigel lumen formation assays, respectively. In addition, the effects of an HIF-1α inhibitor on TSW-containing serum-induced RAEC were also assessed. The effects of TSW-containing serum on the expression of the HIF-1α signaling pathway were evaluated by qRT-PCR and western blot analysis. RESULTS: LC-MS revealed that TSW decoction primarily contained isomaltulose, choline, D-gluconic acid, L-pipecolic acid, hypotaurine, albiflorin, and tryptophan. TSW-containing serum significantly increased the viability, migration, wound healing, and angiogenesis of RAEC in a dose-dependent manner. Furthermore, our results demonstrated that HIF-1α and VEGF expressions were increased in the cells of TSW-containing serum groups, whereas VHL expression was decreased. The effects of TSW-containing serum were reversed by treatment with an HIF-1α inhibitor. CONCLUSION: These results suggested that TSW decoction enhanced angiogenesis by regulating the VHL/HIF-1α/VEGF signaling pathway.

Safflower yellow promotes angiogenesis through p-VHL/ HIF-1α/VEGF signaling pathway in the process of osteogenic differentiation.

OBJECTIVE: Safflower yellow (SY) is an active component ofCarthamus tinctorius L. that is widely used in orthopedics. This study aimed to evaluate the role of SY in angiogenesis and osteogenic differentiation. METHODS: The migration and in vitro angiogenesis of SY (4.5, 9.0, 18 µg/ml)-treated human umbilical vein endothelial cells (HUVEC-12) were assessed by transwell and tube formation assay, respectively. Osteogenic differentiation ability was detected by alkaline phosphatase (ALP) and Alizarin Red S staining. The mRNA and protein expressions of related markers were determined by RT-qPCR and Western blot. RESULTS: The migration and tube formation ability of HUVEC-12 were promoted by SY. Furthermore, SY facilitated the angiogenesis and osteogenic differentiation in the co-culture of HUVEC-12 and BMSCs by increasing hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), Angiopoietin-2 (Ang-2), ALP, runt-related transcription factor 2 (Runx2) and osteopontin-1 (OPN-1) levels. Inhibition of HIF-1α expression by 3-(5-hydroxymethl-2-furyl)-1-benzylindazole (YC-1), restrained SY-induced proliferation, migration and angiogenesis of HUVEC-12 and the increased protein levels of VEGF, Ang-2, ALP, Runx2 and OPN-1. Finally, WD repeat and SOCS box-containing protein-1 (WSB-1)/Von Hippel-Lindau protein (p-VHL) pathway was involved in the beneficial effect of SY. CONCLUSION: SY promotes osteogenic differentiation via enhancing angiogenesis by regulating pVHL/HIF-1α/VEGF signaling pathway.