Chemical and Functional Separation of Pro- and Antiangiogenic Components of Danhong Injection in Ischemic or Tumor Vascular Models

Objective: We aimed to investigate the chemical basis and mechanism of angiogenesis regulation by a multicomponent Chinese medicine Danhong injection (DHI). Methods: A chemical fraction library of DHI was screened and validated for angiogenesis activities by tube formation and migration assays. Mouse ischemic and tumor vascular models were used to verify the angiogenesis effects in vivo. Migration ability of the main monomers of proangiogenic component (PAC) and antiangiogenic component (AAC) in EA.hy926 cells were determined by migration assay. qPCR analyses were performed to access whether the main monomers of PAC or AAC could affect the expression of angiogenesis-related genes in ECs. Western blotting was used to verify the main monomers PAC and AAC effects on CXCR4 protein expression. Results: Two chemically-distinct fractions, including promotion and inhibition of angiogenesis, were identified in DHI. PAC enhanced angiogenesis and improved recovery of ischemic limb perfusion while AAC reduced Lewis lung tumor growth in vivo in VEGFR-2-Luc mice. CA and RA upregulated the expression of TSP1 and downregulated the expression of KDR and PECAM genes. CXCR4 expression was significantly decreased by CA and RA, but increased by PAI, consistent with their differential effects on EC migration. Conclusion: DHI is capable of bi-directional regulation of angiogenesis in a disease-specific manner. The proangiogenesis activity of DHI promotes ischemic vascular injury repair, whereas the anti-angiogenesis activity inhibits tumor growth. The best pro- and anti-angiogenesis activities are composed of unique chemical combinations that differentially regulate angiogenesis-related gene network.