Davallia bilabiata exhibits anti-angiogenic effect with modified MMP-2/TIMP-2 secretion and inhibited VEGF ligand/receptors expression in vascular endothelial cells. (20th January 2017)

• Record Type: Journal Article
• Authors: Liu, Chun-Ting; Bi, Kuo-Wei; Huang, Chao-Chun; Wu, Hsiao-Ting; Ho, Hui-Ya; S. Pang, Jong-Hwei; Huang, Sheng-Teng
• Is Part Of: Journal of ethnopharmacology. Volume 196(2017)
• Page Start: 213
• Page End: 224
• Publication Date: 2017-01-20
• Subjects: D. bilabiata Davallia bilabiata -- MMPs matrix metalloproteinases -- VEGF vascular endothelial growth factor -- VEGFR vascular endothelial growth factor receptor -- CAM chorioallantoic membrane -- TIMP tissue inhibitor of metalloproteinase -- RECK reversion-inducing cysteine-rich protein with kazal motifs -- qRT-PCR real time polymerase chain reaction -- TNF-α tumor necrosis factor-α -- IL-1β interleukin-1β -- CAMs cellular adhesion molecules -- ECM extracellular matrix -- RA rheumatoid arthritis -- D. formosana Davallia formosana -- NF-κB nuclear factor kappa-light-chain-enhancer of activated B cells -- HUVECs human umbilical vascular endothelial cells -- GMP good manufacturing practice -- HPLC high-performance liquid chromatography -- PMA phorbol-12-myristate-13-acetate -- SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis -- PVDF polyvinylidene difluoride -- LDH lactic dehydrogenase; Davallia bilabiata -- Angiogenesis -- Matrix metalloproteinases (MMPs) -- VEGFs -- VEGFRs; Ethnopharmacology -- Periodicals; Pharmacognosy -- Periodicals; Herbs -- Periodicals; Herbs -- Periodicals; Pharmacognosy -- Periodicals; Pharmacognosie -- Périodiques; Herbes -- Périodiques; 615.1
• DOI: 10.1016/j.jep.2016.12.019
• Languages: English
• ISSNs: 0378-8741

Abstract

Abstract: Ethnopharmacological relevance: Davallia bilabiata Hosokawa ( D. bilabiata ), also called GuSuiBu, is popularly used as a substitute for Drynaria fortunei J. Sm for rheumatoid and degenerative arthritis in traditional Chinese medicine. Little is known about the underlying mechanisms of anti-angiogenesis responsible for arthritis in D. bilabiata which needs to be elucidated. Aim of the study: The present study is intended to investigate the anti-angiogenic effect of D. bilabiata associated with the modulation of matrix metalloproteinases (MMPs) and down regulation of vascular endothelial growth factor (VEGF) ligand/receptors both in vivo and in vitro. Materials and methods: We investigated the potential anti-angiogenic effect of D. bilabiata by the in vivo neovascularization of chick chorioallantoic membranes (CAM) assay, and the in vitro migration and matrix-induced tube formation assay using human umbilical vascular endothelial cells (HUVECs). The expressions of MMP-2, TIMP-2, RECK and VEGF/VEGFR were analyzed by real-time RT-PCR or Western blot method. Results: One major compound from water extract of D. bilabiata was identified as Epicatechin 3-O-β-D -allopyranoside. D. bilabiata was confirmed to inhibit in vivo angiogenesis by CAM assay. D. bilabiata also exhibited in vitro anti-angiogenic and anti-regrowth effects as demonstrated by tube formation assay, transwell migration assay and wound healing assay. The mRNA expressions of MMP-2, and MMP-14 were decreased. On the contrary, tissue inhibitor of metalloproteinase-2 (TIMP-2), reversion-inducing cysteine-rich protein with kazal motifs (RECK) were increased by D. bilabiata . The extracellular MMP-2 activity was found to be reduced both in vitro and in vivo by D. bilabiata as determined by gelatin zymography. Results from western blot analysis and ELISA further demonstrated the decrease of MMP-2 and increase of TIMP-2 secretion after D. bilabiata treatment. The gene expressions of VEGF-A, -B, -C, -D and VEGFR-1, -2, -3 were all inhibited by D. bilabiata. Conclusion: We concluded that the anti-angiogenic effect of D. bilabiata was associated with the decreased MMP-2 activity mediated by the upregulation of TIMP-2 and RECK, and the suppression of VEGF/VEGFRs expression. Graphical abstract: Highlights: D. bilabiata inhibited angiogenesis both in vitro and in vivo . The extracellular MMP-2 activity was reduced by D.bilabiata both in vitro and in vivo dose-dependently. D. bilabiata inhibited MMP-2, and MMP-14 and enhanced TIMP-2 and RECK in mRNA level. D. bilabiata induced the dose-dependent decrease of MMP-2 and increase of TIMP-2 secretion. The gene expressions of VEGF-A, -B, -C, -D and VEGFR-1, -2, -3 were all inhibited by D. bilabiata.