Abstract

Background:
Ginseng is a commonly used neutracetical. Intriguingly, existing literature report both wound healing and anti-tumour effects of ginseng extract through opposing activity on the vascular system. To elucidate this perplexity, we merged a chemical fingerprinting approach with a de-constructional study of the effects of pure molecules from ginseng extract on angiogenesis.

Methods and results:
A mass spectrometric compositional analysis of American, Chinese and Korean, and Sanqi ginseng revealed distinct 'sterol ginsenosides' fingerprint, especially in the ratio between a trio1, Rg1, and a dio1, Rb1, the two most prevalent constituents. Using a matrige1 implant model, and reconstituting the extracts using distinct ratios of the two ginsenosides, we demonstrate that the dominance of Rg1 leads to angiogenesis while Rb1 exerts and opposing effect. Rg1 also promoted functional neovascularisation into a polymer scaffold in vivo, and the proliferation of, chemo-invasion, and tubulogenesis by endothelial cells in vitro, an effect mediated through the expression of nitric oxide synthase and the PI3 kinase Akt pathway. In contrast, Rb1 inhibited earliest step in angiogenesis, the chemo-invasion of endothelial cells.

Conclusion:
The current study explains, for the first time, the ambiguity regarding ginseng effects in vascular pathophysiology based on the existence of opposing active principles in the extract. We also unravelled a specio-geographical variation impinging on the compositional fingerprint that may modulate the final phenotype. This emphasises the need for regulations standardizing herbal therapy, currently under the DSHEA. Furthermore, we propose that Rg1 could be a prototype for a novel group of non-peptide molecules that can induce therapeutic angiogenesis such as in wounding healing.

Condensed abstract:
The study is the first to merge a chemical fingerprinting of different ginseng extracts, and identifies novel molecules that exert distinct and sometimes opposing effects on angiogenesis in vivo, and in vitro, explaining the reported ambiguities in the effects of ginseng extracts on the vascular system.

Key words:
Ginsenosides, Angiogenesis, Nitric oxide

Bibliography of Dr. Fan Tai-ping

BSc in Pharmacology, University of London (1979);
PhD in Pharmacology, University of London (1982);
MA, University of Cambridge (1986)

Posts Held:

• Postgraduate Research Assistant (1979-1982), Post-doctoral Research Fellow (1982-1985) and then Bernard Sunley Research Fellow (1985-1986), Department of Pharmacology, Royal College of Surgeons of England; New Blood Lecturer (1986-1988), Lecturer (1988-2001) and then Senior Lecturer/Associate Professor (2001-until retirement) in Pharmacology, Department of Pharmacology, University of Cambridge; Lector in Pharmacology, Trinity College, Cambridge (1992-present)
• University Administration and Committee Membership
• Chairman of Medical & Veterinary Sciences Tripos Part IB and Part II, General Examinations, University of Cambridge (1997-1999);
• Member of Inter-faculty Teaching Committee for Medicine, University of Cambridge (1995- present)
• Honours and Awards, Honorary Member of the Chinese Pharmacological Society (Beijing), elected 1986;
• 1992 FASEB DuP 753 Travel Award to FASEB meeting in Anaheim, California, USA. Apr. 1992;
• Broadhurst Distinguished Lecturer, Schepens Eye Research Institute, Harvard Medical School. (1996);
• Visiting or Honorary Professorships: University of Philippines Manila (1997); School of Chinese Medicine, Hong Kong Baptist University (1998-); Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin, China (1999-); Shanghai Institute of Materia Medica (2002-present); Peking University (2002-present).

Invited lectures at international conferences:

• Invited Speaker: Angiogenesis - Key Principles - Science - Technology - Medicine, St Gallen, Switzerland. Mar, 1991; NATO Advanced Science Institute Conference: Angiogenesis in Health and Disease, Porto Hydra, Greece. Jun. 1991; VII International Symposium on the Biology of Vascular Cells, San Diego, USA. Nov. 1992; 2nd Franz-Volhard Symposium on Angiogenesis, Berlin, Germany. Jun. 1995; 5th Annual Meeting of The German Society of Clinical Pharmacology & Therapeutics, Erfurt, Germany. Nov. 1995; NATO Advanced Science Institute Conference: Angiogenesis - Models, Modulators and Clinical Applications, Rhodes, Greece. Jun. 1997; 3rd World Congress on Inflammation, Tokyo, Japan. Nov. 1997; 3rd International Congress of Pathophysiology, Helsinki, Finland. June/July 1998; 1999 Global Chinese Symposium on Biomaterials and Controlled Release, Taipei, Taiwan. Oct. 1999; 8th Southeast Asia-Western Pacific Regional Meeting of Pharmacologists, Taipei, Taiwan. Nov. 1999; 3rd Asian Conference on Biomaterials and Controlled Release, Taipei, Taiwan. Apr. 2002
• Plenary Speaker: 7th Southeast Asia-Western Pacific Regional Meeting of Pharmacologists, Manila, Philippines. Nov. 1995; First Symposium of the Korean Angiogenesis Research Consortium, Dae Duk Science Town, Korea. Sep. 1999; Japan Angiogenesis Workshop, Tokyo International Conference Center, Tokyo, Apr. 2000; International Conference on Chinese Medicine, Hong Kong Baptist University, Jun. 2001; Institute Pasteur Euro-conference Angiogenesis 1, Paris, March 2001; 2nd World Integrative Medicine Congress, Beijing, China, Sep. 2002; Institute Pasteur Euro-conference Angiogenesis 2, Paris, June 2003; Taipei Conference on Innovation and Biotech Industrial Development, Taipei, Oct. 2003; World Congress of Chinese Medicine, Melbourne, Nov 2003; Tri-Conference: Medicine in the 21st Century ¡V Globalization of Chinese Herbal Medicine, Shanghai, July 2004. Organizer and/or chairman of international conferences
• Cambridge Healthtech Institute Conference "Angiogenesis Antagonists", Bermuda (October 1997); IIR-London Conference "Novel tools and methodologies to promote or inhibit angiogenesis for drug development" (London, December 1998); IIR-New York Conference "Angiogenesis: Therapeutic Applications", Bethesda, MD, USA (February, 1999); IIR-London Conference "ANGIOGENESIS ¡¦99 - Clinical and Pre-clinical Evaluation of Angiogenic Therapeutics " (London, October 1999); First China International Symposium on Angiogenesis & Hematopoiesis: Regulation and Applications in Tianjin, China, September, 1999; Second China International Symposium on Angiogenesis & Hematopoiesis, Tianjin, China, September, 2001; Institute Pasteur Euro-conference Angiogenesis 2, Paris, June 2003; World Congress of Chinese Medicine, Melbourne, Australia, Nov 2003; IUPHAR Congress on Pharmacology, Beijing, July 2006.

Editorial Board etc:

• Special Consultant Editor of "Inflammopharmacology", (1989-present); Editor of "Textbook of Immunopharmacology" with M.M. Dale & J.C. Foreman. 3rd Edition, 1994. Blackwell Scientific Publications; Editor-in-Chief of ANGIOGENESIS (1997-2005); Co-editor of "Immunopharmacology of Traditional Chinese Medicine" 1999 (in Chinese Mianyi Zhongyaoxue, United Press of Beijing Medical University and China Union Medical University); Co-editor of "The New Angiotherapy" with Elise C. Kohn, Humana Press, 2002
• Referee for international journals: "Immunology Letters" (1984-), "Immunology" (1985-) "Biochemical Pharmacology¡¨ (1985-), "Immunopharmacology" (1989-), "British Journal of Cancer" (1989-), "British Journal of Pharmacology" (1993-), "Laboratory Investigation" (1994-), "European Journal of Cancer" (1997-present),"International Journal of Cancer" (1997-), "American Journal of Pathology" (1997-present), "EMBO Journal" (1998-), "Oncology" (1999-present), "FASEB Journal" (1999-present)
• Membership of Learned Societies British Microcirculation Society (1981), British Society for Immunology (1982), International Society for Immunopharmacology (1982), British Pharmacological Society (1983), Honorary Member of Chinese Pharmacological Society (Beijing, 1986), British Association of Cancer Research (1987), Honorary Member of Association of Chinese Traditional Medicine (U.K.) 1994

Publications:

Refereed original papers and submitted manuscripts (invited reviews, book chapters and meeting abstracts on request)

49	Lauder, H., Feniuk, W., Fan, T.-P.D., Humphrey, P.P.A. & Sellers, L.A. (2004). The central role of Src as a determinant of the proliferative outcome of somatostatin sst5 receptor activity in rat aortic vascular smooth muscle cells. Oncogene (submitted)
48	Lauder, H., Alderton, F., Feniuk, W., Fan, T.-P.D., Humphrey, P.P.A. & Sellers, L.A. (2004). Somatostatin sst2 receptor activity inhibits endothelin-1-induced proliferation of vascular smooth muscle cells by abolishing the evoked MAP-kinase response. Circulation Research (submitted)
47	Bevan, D., Gherardi, E., Fan, T.-P.D. & Warn, R. (2004). Diverse and potent activities of HGF/SF in skin wound healing. J Path. (in press).
46	Sengupta, S., Toh, S.-E., Sellers, L.A., Skepper, J., Koolwijk, P., Leung, H.W., Yeung, H.W., Wong, R.N.S., Sasisekharan, R. & Fan, T.-P.D. (2004). Modulating angiogenesis: the yin and the yang in ginseng. Circulation (in press)
45	Bapna, A., Vickerstaffe, E., Warrington, B.H., Ladlow, M., Fan, T.-P.D. & Ley, S.V. (2004). Polymer-assisted, multi-step solution phase synthesis and biological screening of histone deacetylase inhibitors. Org. Biomol. Chem. 2(4):611-20.
44	Sengupta, S., Sellers, L.A., Cindrova, T., Skepper, J., Gherardi, E., Sasisekharan, R, Fan, T.-P.D. (2003). Cyclooxygenase-2-selective nonsteroidal anti-inflammatory drugs inhibit hepatocyte growth factor/scatter factor-induced angiogenesis. Cancer Res. 63(23):8351-9.43 Hull, M.L., Charnock-Jones, D.S., Chan, C.L., Bruner-Tran, K.L., Osteen, K.G., Tom, B.D., Fan, T.-P.D. & Smith, S.K. (2003). Antiangiogenic agents are effective inhibitors of endometriosis. J. Clin. Endocrinol. Metab. 88(6):2889-99.
42	Sengupta, S., Sellers, L.A., Li, R.C., Gherardi, E., Zhao, G., Watson, N., Sasisekharan, R. & Fan, T.-P.D. Targeting of mitogen-activated protein kinases and phosphatidylinositol 3 kinase inhibits hepatocyte growth factor/scatter factor-induced angiogenesis. Circulation, 107(23):2955-61.
41	Sengupta, S., Sellers, L.A., Matheson, H. & Fan, T.-P.D. (2003). Thymidine phoaphorylase induces angiogenesis in vivo and in vitro: an evaluation of possible mechanisms. Br. J. Pharmacol. 139(2):219-31
40	Sengupta, S., Gherardi, E., Sellers, L.A., Wood, J.M., Sasisekharan, R. & Fan, T.-P.D. (2003). Hepatocyte growth factor/scatter factor can induce angiogenesis independently of vascular endothelial growth factor. Arterioscler Thromb Vasc Biol 23(1):69-75
39	Alderton, F., Fan, T.-P.D., Humphrey, P.P.A. (2001). Arachidonic acid mobilisation through recombinant somatostatin receptors: evidence for partial agonism of synthetic peptides. Br. J. Pharmacol., 132: 760-766.
38	Boyle, J.J., Wilson, B., Bicknell, R., Harrower, S., Weissberg, P.L. & Fan, T.-P.D. (2000). Expression of angiogenic factor thymidine phosphorylase and angiogenesis in human atherosclerosis. J. Pathol ., 192, 234-242
37	Alderton, F., Fan, T.-P.D., Schindler, M. & Humphrey, P.P.A. (1999). Rat somatostatin sst2(a) and sst2(b) receptor isoforms mediate opposite effects on cell proliferation. Br. J. Pharmacol., 125: 1630-1633.
36	Smith, E.L.K., Lauder, H., & Fan, T.-P.D., Hiley, C.R. & Hainsworth, A.H. (1998). Vascular endothelial growth factor and endothelin-3 activate cation currents in human umbilical vein endothelial cells. J. Clin. Bas. Cardiol., 1: 52-54
35	Alderton, F., Lauder , H., Feniuk, W., Fan, T.-P.D. & Humphrey, P.P.A. (1998). Comparative effects of somatostatin and angiopeptin on cell proliferation in CHO-K1 cells expressing the human or rat recombinant sst2 or sst5 receptors. Br. J. Pharmacol., 124: 323-330.
34	Lauder, H., Frost, E.E., Hiley, C.R. & Fan, T.-P.D. (1998). Quantification of the repair process involved in the repair of a cell monolayer using an in vitro model of mechanical injury. ANGIOGENESIS, 2: 67-80.
33	Lauder, H., Sellers, L.A., Fan, T.-P.D., Feniuk, W. & Humphrey, P.P.A. (1997). Somatostatin-induced inhibition of the regeneration of rat aortic vascular smooth muscle cells. Br. J. Pharmacol., 122: 663-670.
32	Walsh, D.A., Hu, D.-E., Wharton, J., Catravas, J.D., Blake, D.R. & Fan, T.-P.D. (1997). Sequential development of angiotensin receptors and angiotensin converting enzyme during angiogenesis in the rat subcutaneous sponge granuloma. Br. J. Pharmacol., 120: 1302-1311.
31	Walsh, D.A., Hu, D.-E., Mapp, P.I., Polak, J.M., Blake, D.R. & Fan, T.-P.D. (1996). Innervation and neurokinin receptors during angiogenesis in the rat sponge granuloma. Histochem. J. 28: 759-769.
30	Hori, Y., Hu, D.-E., Yasui, K., Smither, R.L., Gresham, G.A. & Fan, T.-P.D. (1996). Differential effects of angiostatic steroids and dexamethasone on angiogenesis and cytokine levels in rat sponge implants Br. J. Pharmacol., 118: 1584-1591.
29	Hu, D.E., Hiley, C.R. & Fan, T.-P.D.(1996). Comparative studies on the angiogenic activity of vasoactive intestinal peptide, endothelins 1 and 3 and angiotensin II in a rat sponge model. Br. J. Pharmacol. 117: 545-551.
28	Michel, A.D., Chau, N.-M., Fan, T.-P.D., Frost, E.E. & Humphrey, P.P.A.(1995) Evidence that [3H]abmeATP may label an endothelial-derived cell line 5'-nucleotidase with high affinity. Br. J. Pharmacol. 115: 767-774.
27	Lees, V.C., Fan, T.-P.D. & West, D.C. (1995). Angiogenesis in a delayed revascularisation model is accelerated by angiogenic oligosaccharides of hyaluronan. Laboratory Investigation , 73(2): 259-266.
26	Hu, D.E., Hiley, C.R., Smither, R.L., Gresham, G.A. & Fan, T.-P.D. (1995). Correlation of 133Xe clearance, blood flow and histology in the rat sponge model for angiogenesis. Further studies with angiogenic modifiers. Laboratory Investigation, 72(5): 601-610.
25	Moghaddam, A., Zhang, H.-T., Fan, T.-P.D., Hu, D.-E., Lees, V.C., Turley, H., Fox, S.B., Gatter, K.C., Harris, A.L. & Bicknell, R. (1995). Thymidine phosphorylase is angiogenic and promotes tumor growth. Proc. Natl. Acad. Sci., USA. 92: 998-1002.
24	Hu, D.E. & Fan, T.-P.D. (1995). Suppression of VEGF-induced angiogenesis by the protein tyrosine kinase inhibitor, lavendustin A. Br. J. Pharmacol. 114: 262-268
23	Hu, D.E. & Fan, T.-P.D. (1995). Protein kinase C inhibitor calphostin C prevents cytokine-induced angiogenesis in vivo. Inflammation, 19 (1): 39-54
22	Lees, V.C. & Fan, T.-P.D. (1994). A freeze-injured skin graft model for the quantitative study of basic fibroblast growth factor and other promoters of angiogenesis in wound healing. Br. J. Plast. Surg., 47: 349-359.
21	Braddock, P.S., Hu, D.-E., Fan, T.-P.D., Stratford, I.J., Harris, A.L. & Bicknell, R. (1994). A structure-activity analysis of antagonism of the growth factor and angiogenic activity of basic fibroblast growth factor by suramin and related polyanions. Br. J. Cancer, 69: 890-898.
20	Hu, D.E., Hori, Y., Presta, M., Gresham, G.A. & Fan, T.-P.D. (1994). Inhibition of angiogenesis by IL-1 receptor antagonist and selected cytokine antibodies in rats. Inflammation, 18: 45-58.
19	Wren, A.D., Hiley, C.R. & Fan, T.-P.D. (1993). Endothelin-3 mediated proliferation in wounded human umbilical vein endothelial cells. Biochem. Biophys. Res. Commun., 196: 369-375.
18	Fan, T.-P.D., Hu, D.E., Guard, S., Gresham, G.A. & Watling, K.J. (1993). Stimulation of angiogenesis by substance P and interleukin-1 in rats and its inhibition by NK1 and interleukin-1 receptor antagonists. Br. J. Pharmacol., 110: 43-49
17	Hu, D.E. & Fan, T.-P.D. (1993) Leu8]des-Arg9-bradykinin inhibits the angiogenic effect of bradykinin and interleukin-1 in rats. Br. J. Pharmacol., 109: 14-17
16	Hu, D.E., Hori, Y. & Fan, T.-P.D. (1993) Interleukin 8 stimulates angiogenesis in rats. Inflammation, 17: 135-143
15	Dinh Xuan, A.T., Fan, T.-P.D., Higenbottam, T.W. & Wallwork, J. (1990) Cyclosporine in vitro reduces endothelium-dependent relaxation to acetylcholine, but does not affect relaxation to nitrovasodilators. Transplantation Proceedings 22: 1723-1725
14	Lo, W.W.Y. & Fan, T.-P.D. (1987b) Bradykinin stimulates inositol phosphate accumulation in cultured human umbilical vein endothelial cells. Med. Sci. Res. 15: 1325
13	Lo, W.W.Y. & Fan, T.-P.D. (1987a) Histamine stimulates inositol phosphate accumulation via the H1-receptor in cultured human endothelial cells. Biochem. Biophys. Res. Commun. 148: 47-53
12	Andrade, S.P., Fan, T.-P.D. & Lewis, G.P. (1987) Quantitative in vivo studies on angiogenesis in a rat sponge model. Br. J. Exp. Path. 68: 755-766
11	Fan, T.-P.D., Cox, J.H. & Chisholm, P.M. (1987) Mechanism of action of cyclosporine in preventing cardiac allograft rejection. II. Graft tissue levels of prostacyclin and thromboxane. Transplantation 43: 343-345
10	Kalmar, L., Fan, T.-P.D. & Lewis, G.P. (1986) Zymosan-air-pouch model of acute inflammation. Adv. Pharmacol. Res. Pract. 3: 527-531
9	Kalmar, L., Fan, T.-P.D., Lewis, G.P. & Feher, J. (1986) Formation of prostaglandin E2, prostacyclin and leukotriene B4 and their effect during the mixed lymphocyte reaction of rat spleen cells. Kiserl. Orvostud. 38 (5): 449-456
8	Kalmar, L., Fan, T.-P.D., Lewis, G.P., Kadar, J. & Feher, J. (1986) Effect of eicosanoids (PGE2, LTB4) and lymphokine on granulocyte accumulation in vivo. Kiserl. Orvostud. 38 (1): 38-43
7	Cole, O.F., Fan, T.-P.D. & Lewis, G.P. (1986b) Release of eicosanoids from cultured rat aortic endothelial cells; studies with arachidonic acid and calcium ionophore A23187. Cell Biology International Reports 10: 407-413
6	Cole, O.F., Fan, T.-P.D. & Lewis, G.P. (1986a) Isolation, characterisation, growth and culture of endothelial cells from the rat aorta. Cell Biology International Reports 10: 399-405
5	Kalmar, L., Fan, T.-P.D., Lewis, G.P. & Feher, J. (1985b) Dynamics of a model of acute inflammation induced by zymosan in a subcutaneous blister. Kiserl. Orvostud. 37 (6): 581-587
4	Kalmar, L., Fan, T.-P.D., Lewis, G.P. & Feher, J. (1985a) Mechanism of the elimination of the eicosanoid effect in acute inflammation. Kiserl. Orvostud. 37 (5): 503-511
3	Fan, T.-P.D. & Lewis, G.P. (1985) Mechanism of cyclosporin-A-induced inhibition of prostacyclin synthesis by macrophages. Prostaglandins 30: 735-743
2	Fan, T.-P.D. & Lewis, G.P. (1984) Effect of cyclosporin A and inhibitors of arachidonic acid metabolism on blood flow and cyclo-oxygenase products in rat skin allografts. Br. J. Pharmacol. 81: 261-271
1	Fan, T.-P.D. & Lewis, G.P. (1982) Blood flow, histamine content and histidine decarboxylase activity in rat skin grafts and their modification by cyclosporin A. Br. J. Pharmacol. 76: 491-497