FOLLOWUS
Pro-angiogenic activity of notoginsenoside R1 in human umbilical vein endothelial cells in vitro and in a chemical-induced blood vessel loss model of zebrafish in vivo
- Chinese Journal of Integrative Medicine Vol. 22, Issue 6, Pages: 420-429(2016)
- Affiliations：
1. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao,China
2. School of Chinese Medicine, The Chinese University of Hong Kong,Hong Kong SAR,China
3. Laboratory of Cardiovascular Diseases, Xiyuan Hospital China Heart Institute of Chinese Medicine, China Academy of Chinese Medical Sciences,Beijing,China
- Author bio：
- Funds：Supported by grants from the Overseas and Hong Kong, Macau Young Scholars Collaborative Research Fund by the National Natural Science Foundation of China (No. 81328025) and the Science and Technology Development Fund of Macau SAR (Ref. No. 014/2011/A1), Research Committee, University of Macau;These authors contributed equally to this work
DOI：10.1007/s11655-014-1954-8
CLC：Published：2016，
Published Online：22 December 2014，
Scan for full text
Yang, Br., Hong, Sj., Lee, S.MY. et al. Pro-angiogenic activity of notoginsenoside R1 in human umbilical vein endothelial cells in vitro and in a chemical-induced blood vessel loss model of zebrafish in vivo, ., Chin. J. Integr. Med. 22, 420–429 (2016). https://doi.org/10.1007/s11655-014-1954-8
Bin-rui Yang, Si-jia Hong, Simon Ming-Yuen Lee, et al. Pro-angiogenic activity of notoginsenoside R1 in human umbilical vein endothelial cells in vitro and in a chemical-induced blood vessel loss model of zebrafish in vivo. [J]. Chinese Journal of Integrative Medicine 22(6):420-429(2016)
Yang, Br., Hong, Sj., Lee, S.MY. et al. Pro-angiogenic activity of notoginsenoside R1 in human umbilical vein endothelial cells in vitro and in a chemical-induced blood vessel loss model of zebrafish in vivo, ., Chin. J. Integr. Med. 22, 420–429 (2016). https://doi.org/10.1007/s11655-014-1954-8 DOI：
Bin-rui Yang, Si-jia Hong, Simon Ming-Yuen Lee, et al. Pro-angiogenic activity of notoginsenoside R1 in human umbilical vein endothelial cells in vitro and in a chemical-induced blood vessel loss model of zebrafish in vivo. [J]. Chinese Journal of Integrative Medicine 22(6):420-429(2016) DOI： 10.1007/s11655-014-1954-8.
摘要
This study aimed at investigating whether notoginsenoside R1 (R1)
a unique saponin found in Panax notoginseng could promote angiogenic activity on human umbilical vein endothelial cells (HUVECs) and elucidate their potential molecular mechanisms. In addition
vascular restorative activities of R1 was assessed in a chemically-induced blood vessel loss model in zebrafish. The in vitro angiogenic effect of R1 was compared with other previously reported angiogenic saponins Rg1 and Re. The HUVECs proliferation in the presence of R1 was determined by cell proliferation kit II (XTT) assay. R1
Rg1 and Re-induced HUVECs invasion across polycarbonate membrane was stained with Hoechst-33342 and quantified microscopically. Tube formation assay using matrigelcoated wells was performed to evaluate the pro-angiogenic actions of R1. In order to understand the mechanism underlying the pro-angiogenic effect
various pathway inhibitors such as SU5416
wortmannin (wort) or L-Nω-nitro- L-arginine methyl ester hydrochloride (L-NAME)
SH-6 were used to probe the possible involvement of signaling pathway in the R1 mediated HUVECs proliferation. In in vivo assays
zebrafish embryos at 21 hpf were pre-treated with vascular endothelial growth factor (VEGF) receptor kinase inhibitor II (VRI) for 3 h only and subsequently post-treated with R1 for 48 h
respectively. The intersegmental vessels (ISVs) in zebrafish were assessed for the restorative effect of R1 on defective blood vessels. R1 could stimulate the proliferation of HUVECs. In the chemoinvasion assay
R1 significantly increased the number of cross-membrane HUVECs. In addition
R1 markedly enhanced the tube formation ability of HUVECs. The proliferative effects of these saponins on HUVECs were effectively blocked by the addition of SU5416 (a VEGF-KDR/Flk-1 inhibitor). Similarly
pre-treatment with wort [a phosphatidylinositol 3-kinase (PI3K)-kinase inhibitor]
L-NAME [an endothelial nitric oxide synthase (eNOS) inhibitor] or SH-6 (an Akt pathway inhibitor) significantly abrogated the R1 induced proliferation of HUVECs. In chemicallyinduced blood vessel loss model in zebrafish
R1 significantly rescue the damaged ISVs. R1
similar to Rg1 and Re
had been showed pro-angiogenic action
possibly via the activation of the VEGF-KDR/Flk-1 and PI3K-Akt-eNOS signaling pathways. Our findings also shed light on intriguing pro-angiogenic effect of R1 under deficient angiogenesis condition in a pharmacologic-induced blood vessels loss model in zebrafish. The present study in vivo and in vitro provided scientific evidence to explain the ethnomedical use of Panax notoginseng in the treatment of cardiovascular diseases
traumatic injuries and wound healing.
Abstract
This study aimed at investigating whether notoginsenoside R1 (R1)
a unique saponin found in Panax notoginseng could promote angiogenic activity on human umbilical vein endothelial cells (HUVECs) and elucidate their potential molecular mechanisms. In addition
vascular restorative activities of R1 was assessed in a chemically-induced blood vessel loss model in zebrafish. The in vitro angiogenic effect of R1 was compared with other previously reported angiogenic saponins Rg1 and Re. The HUVECs proliferation in the presence of R1 was determined by cell proliferation kit II (XTT) assay. R1
Rg1 and Re-induced HUVECs invasion across polycarbonate membrane was stained with Hoechst-33342 and quantified microscopically. Tube formation assay using matrigelcoated wells was performed to evaluate the pro-angiogenic actions of R1. In order to understand the mechanism underlying the pro-angiogenic effect
various pathway inhibitors such as SU5416
wortmannin (wort) or L-Nω-nitro- L-arginine methyl ester hydrochloride (L-NAME)
SH-6 were used to probe the possible involvement of signaling pathway in the R1 mediated HUVECs proliferation. In in vivo assays
zebrafish embryos at 21 hpf were pre-treated with vascular endothelial growth factor (VEGF) receptor kinase inhibitor II (VRI) for 3 h only and subsequently post-treated with R1 for 48 h
respectively. The intersegmental vessels (ISVs) in zebrafish were assessed for the restorative effect of R1 on defective blood vessels. R1 could stimulate the proliferation of HUVECs. In the chemoinvasion assay
R1 significantly increased the number of cross-membrane HUVECs. In addition
R1 markedly enhanced the tube formation ability of HUVECs. The proliferative effects of these saponins on HUVECs were effectively blocked by the addition of SU5416 (a VEGF-KDR/Flk-1 inhibitor). Similarly
pre-treatment with wort [a phosphatidylinositol 3-kinase (PI3K)-kinase inhibitor]
L-NAME [an endothelial nitric oxide synthase (eNOS) inhibitor] or SH-6 (an Akt pathway inhibitor) significantly abrogated the R1 induced proliferation of HUVECs. In chemicallyinduced blood vessel loss model in zebrafish
R1 significantly rescue the damaged ISVs. R1
similar to Rg1 and Re
had been showed pro-angiogenic action
possibly via the activation of the VEGF-KDR/Flk-1 and PI3K-Akt-eNOS signaling pathways. Our findings also shed light on intriguing pro-angiogenic effect of R1 under deficient angiogenesis condition in a pharmacologic-induced blood vessels loss model in zebrafish. The present study in vivo and in vitro provided scientific evidence to explain the ethnomedical use of Panax notoginseng in the treatment of cardiovascular diseases
traumatic injuries and wound healing.
关键词
notoginsenoside R1ginsenoside Rg1ginsenoside ReHuman Umbilical Vein Endothelial Cellzebrafishangiogenesis
Keywords
notoginsenoside R1ginsenoside Rg1ginsenoside ReHuman Umbilical Vein Endothelial Cellzebrafishangiogenesis
references
Arima S, Nishiyama K, Ko T, Arima Y, Hakozaki Y, Sugihara K, et al. Angiogenic morphogenesis driven by dynamic and heterogeneous collective endothelial cell movement. Development 2011;138:4763–4776.
Chatterjee S, Chapman KE, Fisher AB. Lung ischemia: a model for endothelial mechanotransduction. Cell Biochem Biophys 2008;52:125–138.
Chung BH, Lee JJ, Kim JD, Jeoung D, Lee H, Choe J, et al. Angiogenic activity of sesamin through the activation of multiple signal pathways. Biochem Biophys Res Commun 2010;391:254–260.
Ferrara N. Vascular endothelial growth factor: basic science and clinical progress. Endocr Rev 2004;25:581–611.
Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R, Zeiher AM. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 1999;399:601–605.
Liu J, Wang Y, Qiu L, Yu Y, Wang C. Saponins of Panax notoginseng: chemistry, cellular targets and therapeutic opportunities in cardiovascular diseases. Expert Opin Investig Drugs 2014;23:523–539.
Wan JB, Lee SM, Wang JD, Wang N, He CW, Wang YT, et al. Panax notoginseng reduces atherosclerotic lesions in ApoE-deficient mice and inhibits TNF-alpha-induced endothelial adhesion molecule expression and monocyte adhesion. J Agric Food Chem 2009;57:6692–6697.
Wang N, Wan JB, Chan SW, Deng YH, Yu N, Zhang QW, et al. Comparative study on saponin fractions from Panax notoginseng inhibiting inflammation-induced endothelial adhesion molecule expression and monocyte adhesion. Chin Med 2011;6:37.
Gao B, Huang L, Liu H, Wu H, Zhang E, Yang L, et al. Platelet P2Y(1)(2) receptors are involved in the haemostatic effect of notoginsenoside Ft1, a saponin isolated from Panax notoginseng. Br J Pharmacol 2014;171:214–223.
Uzayisenga R, Ayeka PA, Wang Y. Anti-diabetic potential of Panax Notoginseng Saponins (PNS): a review. Phytother Res 2014;28:510–516.
Ning N, Dang X, Bai C, Zhang C, Wang K. Panax notoginsenoside produces neuroprotective effects in rat model of acute spinal cord ischemia-reperfusion injury. J Ethnopharmacol 2012;139:504–512.
Usami Y, Liu YN, Lin AS, Shibano M, Akiyama T, Itokawa H, et al. Antitumor agents. 261. 20(S)-protopanaxadiol and 20(S)-protopanaxatriol as antiangiogenic agents and total assignment of H-1 NMR spectrao. J Natural Products 2008;71:478–481.
Wan JB, Yang FQ, Li SP, Wang YT, Cui XM. Chemical characteristics for different parts of Panax notoginseng using pressurized liquid extraction and HPLC-ELSD. J Pharm Biomed Anal 2006;41:1596–1601.
Yu LC, Chen SC, Chang WC, Huang YC, Lin KM, Lai PH, et al. Stability of angiogenic agents, ginsenoside Rg1 and Re, isolated from Panax ginseng: in vitro and in vivo studies. Int J Pharm 2007;328:168–176.
Lee YJ, Chung E, Lee KY, Lee YH, Huh B, Lee SK. Ginsenoside-Rg1, one of the major active molecules from Panax ginseng, is a functional ligand of glucocorticoid receptor. Mol Cell Endocrinol 1997;133:135–140.
Leung KW, Cheng YK, Mak NK, Chan KK, Fan TP, Wong RN. Signaling pathway of ginsenoside-Rg1 leading to nitric oxide production in endothelial cells. FEBS Lett 2006;580:3211–3216.
Ling CQ, Yong L, Zhu XY, Chen Z, Min L. Ginsenosides may reverse the dexamethasone-induced down-regulation of glucocorticold receptor. Gener Compar Endocrinol 2005;140:203–209.
Sengupta S, Toh SA, Sellers LA, Skepper JN, Koolwijk P, Leung HW, et al. Modulating angiogenesis: the yin and the yang in ginseng. Circulation 2004;110:1219–1225.
Furukawa T, Bai CX, Kaihara A, Ozaki E, Kawano T, Nakaya Y, et al. Ginsenoside Re, a main phytosterol of Panax ginseng, activates cardiac potassium channels via a nongenomic pathway of sex hormones. Mol Pharmacol 2006;70:1916–1924.
Zhang HS, Wang SQ. Notoginsenoside R1 from Panax notoginseng inhibits TNF-alpha-induced PAI-1 production in human aortic smooth muscle cells. Vascul Pharmacol 2006;44:224–230.
Sun B, Xiao J, Sun XB, Wu Y. Notoginsenoside R1 attenuates cardiac dysfunction in endotoxemic mice: an insight into oestrogen receptor activation and PI3K/Akt signalling. Br J Pharmacol 2013;168:1758–1770.
Meng X, Sun G, Ye J, Xu H, Wang H, Sun X. Notoginsenoside R1-mediated neuroprotection involves estrogen receptor-dependent crosstalk between Akt and ERK1/2 pathways: a novel mechanism of Nrf2/ARE signaling activation. Free Radic Res 2014;48:445–460.
Liu WJ, Tang HT, Jia YT, Ma B, Fu JF, Wang Y, et al. Notoginsenoside R1 attenuates renal ischemia-reperfusion injury in rats. Shock 2010;34:314–320.
Lam HW, Lin HC, Lao SC, Gao JL, Hong SJ, Leong CW, et al. The angiogenic effects of Angelica sinensis extract on HUVEC in vitro and zebrafish in vivo. J Cell Biochem 2008;103:195–211.
Tanimoto T, Jin ZG, Berk BC. Transactivation of vascular endothelial growth factor (VEGF) receptor Flk-1/KDR is involved in sphingosine 1-phosphate-stimulated phosphorylation of Akt and endothelial nitric-oxide synthase (eNOS). J Biol Chem 2002;277:42997–43001.
Isogai S, Lawson ND, Torrealday S, Horiguchi M, Weinstein BM. Angiogenic network formation in the developing vertebrate trunk. Development 2003;130:5281–5290.
Sun HX, Chen Y, Ye Y. Ginsenoside Re and notoginsenoside R1: immunologic adjuvants with low haemolytic effect. Chem Biodivers 2006;3:718–726.
Milkiewicz M, Hudlicka O, Brown MD, Silgram H. Nitric oxide, VEGF, and VEGFR-2: interactions in activity-induced angiogenesis in rat skeletal muscle. Am J Physiol Heart Circ Physiol 2005;289:H336–H343.
Bocci G, Danesi R, Marangoni G, Fioravanti A, Boggi U, Esposito I, et al. Antiangiogenic versus cytotoxic therapeutic approaches to human pancreas cancer: an experimental study with a vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor and gemcitabine. Eur J Pharmacol 2004;498:9–18.
Kiselyov A, Balakin KV, Tkachenko SE. VEGF/VEGFR signalling as a target for inhibiting angiogenesis. Expert Opinion Investig Drugs 2007;16:83–107.
Gille H, Kowalski J, Li B, LeCouter J, Moffat B, Zioncheck TF, et al. Analysis of biological effects and signaling properties of Flt-1 (VEGFR-1) and KDR (VEGFR-2). A reassessment using novel receptor-specific vascular endothelial growth factor mutants. J Biol Chem 2001;276:3222–3230.
Bussolati B, Dunk C, Grohman M, Kontos CD, Mason J, Ahmed A. Vascular endothelial growth factor receptor-1 modulates vascular endothelial growth factor-mediated angiogenesis via nitric oxide. Am J Pathol 2001;159:993–1008.
Bocci G, Man S, Green SK, Francia G, Ebos JML, du Manoir JM, et al. Increased plasma vascular endothelial growth factor (VEGF) as a surrogate marker for optimal therapeutic dosing of VEGF receptor-2 monoclonal antibodies. Cancer Res 2004;64:6616–6625.
Leung KW, Pon YL, Wong RNS, Wong AST. Ginsenoside-Rg1 induces vascular endothelial growth factor expression through the glucocorticoid receptor-related phosphatidylinositol 3-kinase/Akt and beta-catenin/T-cell factor-dependent pathway in human endothelial cells. J Biol Chemi 2006;281:36280–36288.
Miscevic F, Rotstein O, Wen XY. Advances in zebrafish high content and high throughput technologies. Comb Chem High Throughput Screen 2012;15:515–521.
Hung MW, Zhang ZJ, Li S, Lei B, Yuan S, Cui GZ, et al. From omics to drug metabolism and high content screen of natural product in zebrafish: a new model for discovery of neuroactive compound. Evid Based Complement Alternat Med 2012;2012:605303.
Limsuwanchote S, Wungsintaweekul J, Yusakul G, Han JY, Sasaki-Tabata K, Tanaka H, et al. Preparation of a monoclonal antibody against notoginsenoside R1, a distinctive saponin from Panax notoginseng, and its application to indirect competitive ELISA. Planta Med 2014;80:337–342.
0
Views
0
Downloads
3
CSCD
- Meta-XMLDownload
- BibTeXDownload
- EndnoteDownload
- RefMan Download
- NoteExpressDownload
- NoteFirstDownload
Publicity Resources
Related Articles
Related Author
Related Institution
- Address：1 Caochang, xiyuan, Beijing, People's Republic of China Postal code：100091
- Tel：010-62886827, 62877592 Fax：010-62874291
- Technical support is provided by Beijing Founder electronics co., LTD 京ICP备05067934号-1
京公网安备11010802024621 - It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
- Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.
