Drynaria fortunei Promoted Angiogenesis Associated With Modified MMP-2/TIMP-2 Balance and Activation of VEGF Ligand/Receptors Expression

Abstract

Background and Purpose: Drynaria fortunei J. Sm (D. fortunei), known as Gu-Sui-Bu, is used in traditional Chinese medicine to treat common injuries, including bone fractures and bruising. The specific functional mechanisms of the angiogenic and endothelial cell migration properties of D. fortunei are currently unclear. Thus, the purpose of this study is to validate the potential angiogenic and cellular migration properties and related mechanisms by D. fortunei both in vivo and in vitro.Experimental Approach: The present study investigates, both in vivo and in vitro, the wound healing effects of D. fortunei as associated with angiogenesis, specifically by the modulation of matrix metalloproteinases (MMPs) and upregulation of vascular endothelial growth factor (VEGF) ligand/receptors. In order to determine the potential angiogenic effects of D. fortunei, in vivo neovascularization of chick chorioallantoic membranes (CAMs) assay, and directed in vivo angiogenesis assay (DIVVA) were performed, while in vitro scratch wound healing, migration, and matrix-induced tube formation assays were performed by using human umbilical vascular endothelial cells (HUVECs). Furthermore, we used qPCR to analyze the gene expressions and Western blot to observe protein expressions of MMP-2, MMP-14, TIMP-2, RECK, and VEGF/VEGFRs.Results: This study identified five major compounds from the water extract of D. fortunei: protocatechuic acid, caffeic acid 4-O-β-D-glucopyranoside, 5,7-dihydroxychromone-7-O-rutinoside, neoeriocitrin, and naringin. D. fortunei was confirmed to activate in vivo angiogenesis by CAM and DIVVA assays. D. fortunei further exhibited in vitro angiogenic effects associated with cell migration, as demonstrated by the tube formation assay, transwell migration assay, and scratch wound healing assay. The extracellular MMP-2 activity was found to be dose-dependently augmented both in vitro and in vivo by D. fortunei. The mRNA and protein expressions of MMP-2, and MMP-14 were increased; while the tissue inhibitor metalloproteinase-2 (TIMP-2), and reversion-inducing cysteine-rich protein with kazal motifs (RECK) were both decreased. Furthermore, D. fortunei activated the gene and protein expressions of VEGF-A, -B, and VEGFR-2, -3.Conclusion: D. fortunei increased MMP-2 activity, thereby stimulating angiogenesis and cell migration, both in vivo and in vitro, as a result of MMP-2 and TIMP-2 balance modulation and the activation of VEGF/VEGFRs expression.

Bioinformatics and Molecular Insights to Anti-Metastasis Activity of Triethylene Glycol Derivatives

“…There are 28 identified subtypes of MMPs; MMPs can be categorized into collagenases (MMP-1, -8 and -13), stromelysins (MMP-3 and -10), gelatinases (MMP-2 and -9) and matrilysins (MMP-7 and -26) [ 21 ]. MMP-2 is one of the proteins produced directly downstream to VEGF [ 22 ]. Structurally, these have four major domains–(i) the zinc-interacting cysteine switch containing pro-peptide responsible to keep the protein in an inactivated state, (ii) the zinc-ion binding motif harboring catalytic domain, (iii) the hinge region responsible for enzyme’s stability and (iv) the hemopexin-like C-terminal domain organized into a four-bladed -propeller structure and responsible for protein interaction and substrate stability.…”

Treatment of tibial dyschondroplasia with traditional Chinese medicines: “Lesson and future directions”

“…Thiram can change the differentiation of abnormal chondrocytes by altering the expression of the HIF-1α/VEGF pathway of chondrocytes. Previously, Drynaria fortunei promoted angiogenesis associated with modified MMP-2/TIMP-2 balance ( Mehmood et al., 2018a ; Huang et al., 2018 ). Icariin can significantly improve abnormal angiogenesis in the GP of TD and promote vascular recovery ( Zhang et al., 2018c ).…”

Chemometric Analyses for the Characterization of Raw and Stir-Frying Processed Drynariae Rhizoma Based on HPLC Fingerprints

“…Neoeriocitrin also significantly increased proliferation of UMR 106 cells and exhibited effects on proliferation and osteogenic cell differentiation in MC3T3-E1 [ 16 , 17 ]. Furthermore, phenolic acids in DR increased MMP-2 activity and stimulated angiogenesis and cell migration in vivo and in vitro [ 18 ]. In addition, drynachromoside A exhibited the biochemical effects on the proliferation of MC3T3-E1 cells [ 19 ].…”