Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Inhibitory effects of total saponin from Korean red ginseng via vasodilator-stimulated phosphoprotein-Ser157 phosphorylation on thrombin-induced platelet aggregation

  • Lee, Dong-Ha (Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering and Regional Research Center, Inje University) ;
  • Cho, Hyun-Jeong (Department of Biomedical Laboratory Science, College of Medical Science, Konyang University) ;
  • Kim, Hyun-Hong (Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering and Regional Research Center, Inje University) ;
  • Rhee, Man Hee (Laboratory of Veterinary Physiology & Signaling, College of Veterinary Medicine, Kyungpook National University) ;
  • Ryu, Jin-Hyeob (Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering and Regional Research Center, Inje University) ;
  • Park, Hwa-Jin (Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering and Regional Research Center, Inje University)
  • Received : 2012.09.04
  • Accepted : 2012.12.05
  • Published : 2013.04.15
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Abstract

In this study, we have investigated the effects of total saponin from Korean red ginseng (TSKRG) on thrombin-induced platelet aggregation. TSKRG dose-dependently inhibited thrombin-induced platelet aggregation with $IC_{50}$ value of about 81.1 ${\mu}g/mL$. In addition, TSKRG dose-dependently decreased thrombin-elevated the level of cytosolic-free $Ca^{2+}$ ($[Ca^{2+}]_i$), one of aggregation-inducing molecules. Of two $Ca^{2+}$-antagonistic cyclic nucleotides as aggregation-inhibiting molecules, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), TSKRG significantly dose-dependently elevated intracellular level of cAMP, but not cGMP. In addition, TSKRG dose-dependently inhibited thrombin-elevated adenosine triphosphate (ATP) release from platelets. These results suggest that the suppression of $[Ca^{2+}]_i$ elevation, and of ATP release by TSKRG are associated with upregulation of cAMP. TSKRG elevated the phosphorylation of vasodilator-stimulated phosphoprotein (VASP)-$Ser^{157}$, a cAMP-dependent protein kinase (A-kinase) substrate, but not the phosphorylation of VASP-$Ser^{239}$, a cGMP-dependent protein kinase substrate, in thrombin-activated platelets. We demonstrate that TSKRG involves in increase of cAMP level and subsequent elevation of VASP-$Ser^{157}$ phosphorylation through A-kinase activation to inhibit $[Ca^{2+}]_i$ mobilization and ATP release in thrombin-induced platelet aggregation. These results strongly indicate that TSKRG is a beneficial herbal substance elevating cAMP level in thrombin-platelet interaction, which may result in preventing of platelet aggregation-mediated thrombotic diseases.

Keywords

References

  1. Schwartz SM, Heimark RL, Majesky MW. Developmental mechanisms underlying pathology of arteries. Physiol Rev 1990;70:1177-1209. https://doi.org/10.1152/physrev.1990.70.4.1177
  2. Berridge MJ, Irvine RF. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature 1984;312:315-321. https://doi.org/10.1038/312315a0
  3. Jennings LK. Role of platelets in atherothrombosis. Am J Cardiol 2009;103:4A-10A.
  4. Pasqui AL, Capecchi PL, Ceccatelli L, Mazza S, Gistri A, Laghi Pasini F, Di Perri T. Nitroprusside in vitro inhibits platelet aggregation and intracellular calcium translocation. Effect of haemoglobin. Thromb Res 1991;61:113-122.
  5. Nishikawa M, Tanaka T, Hidaka H. $Ca^{2+}$-calmodulin-dependent phosphorylation and platelet secretion. Nature 1980;287:863-865. https://doi.org/10.1038/287863a0
  6. Halbrugge M, Walter U. Purification of a vasodilator-regulated phosphoprotein from human platelets. Eur J Biochem 1989;185:41-50. https://doi.org/10.1111/j.1432-1033.1989.tb15079.x
  7. Halbrugge M, Friedrich C, Eigenthaler M, Schanzenbacher P, Walter U. Stoichiometric and reversible phosphorylation of a 46-kDa protein in human platelets in response to cGMP- and cAMP-elevating vasodilators. J Biol Chem 1990;265:3088-3093.
  8. Butt E, Abel K, Krieger M, Palm D, Hoppe V, Hoppe J, Walter U. cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. J Biol Chem 1994;269:14509-14517.
  9. Homer KL, Wanstall JC. Inhibition of rat platelet aggregation by the diazeniumdiolate nitric oxide donor MAHMA NONOate. Br J Pharmacol 2002;137:1071-1081. https://doi.org/10.1038/sj.bjp.0704971
  10. Park WH, Kim HK, Nam KS, Shon YH, Jeon BH, Moon SK, Kim MG, Kim CH. Inhibitory effect of GBH on platelet aggregation through inhibition of intracellular Ca2+ mobilization in activated human platelets. Life Sci 2004;75:3063-3076. https://doi.org/10.1016/j.lfs.2004.07.010
  11. Laurent V, Loisel TP, Harbeck B, Wehman A, Grobe L, Jockusch BM, Wehland J, Gertler FB, Carlier MF. Role of proteins of the Ena/VASP family in actin-based motility of Listeria monocytogenes. J Cell Biol 1999;144:1245-1258. https://doi.org/10.1083/jcb.144.6.1245
  12. Smolenski A, Bachmann C, Reinhard K, Honig-Liedl P, Jarchau T, Hoschuetzky H, Walter U. Analysis and regulation of vasodilator-stimulated phosphoprotein serine 239 phosphorylation in vitro and in intact cells using a phosphospecific monoclonal antibody. J Biol Chem 1998;273:20029-20035. https://doi.org/10.1074/jbc.273.32.20029
  13. Horstrup K, Jablonka B, Honig-Liedl P, Just M, Kochsiek K, Walter U. Phosphorylation of focal adhesion vasodilator-stimulated phosphoprotein at Ser157 in intact human platelets correlates with fibrinogen receptor inhibition. Eur J Biochem 1994;225:21-27. https://doi.org/10.1111/j.1432-1033.1994.00021.x
  14. Ernst E. Panax ginseng: an overview of the clinical evidence. J Ginseng Res 2010;34:259-263. https://doi.org/10.5142/jgr.2010.34.4.259
  15. Kim SK, Park JH. Trends in ginseng research in 2010. J Ginseng Res 2011;35:389-398. https://doi.org/10.5142/jgr.2011.35.4.389
  16. Chung IM, Lim JW, Pyun WB, Kim H. Korean red ginseng improves vascular stiffness in patients with coronary artery disease. J Ginseng Res 2010;34:212-218. https://doi.org/10.5142/jgr.2010.34.3.212
  17. Wee JJ, Kim YS, Kyung JS, Song YB, Do JH, Kim DC, Lee SD. Identification of anticoagulant components in Korean red ginseng. J Ginseng Res 2010;34:355-362. https://doi.org/10.5142/jgr.2010.34.4.355
  18. Jung YH, Park KY, Jeon JH, Kwak YS, Song YB, Wee JJ, Rhee MH, Kim TW. Red ginseng saponin fraction A isolated from Korean red ginseng by ultrafiltration on the porcine coronary artery. J Ginseng Res 2011;35:325-330. https://doi.org/10.5142/jgr.2011.35.3.325
  19. Hwang SY, Son DJ, Kim IW, Kim DM, Sohn SH, Lee JJ, Kim SK. Korean red ginseng attenuates hypercholesterolemia-enhanced platelet aggregation through suppression of diacylglycerol liberation in high-cholesterol-diet-fed rabbits. Phytother Res 2008;22:778-783. https://doi.org/10.1002/ptr.2363
  20. Lee DH, Cho HJ, Kang HY, Rhee MH, Park HJ. Total saponin from Korean red ginseng inhibits thromboxane A2 production associated microsomal enzyme activity in platelets. J Ginseng Res 2012;36:40-46. https://doi.org/10.5142/jgr.2012.36.1.40
  21. Schaeffer J, Blaustein MP. Platelet free calcium concentrations measured with fura-2 are influenced by the transmembrane sodium gradient. Cell Calcium 1989;10:101-113. https://doi.org/10.1016/0143-4160(89)90050-X
  22. Kim SN, Ha YW, Shin H, Son SH, Wu SJ, Kim YS. Simultaneous quantification of 14 ginsenosides in Panax ginseng C.A. Meyer (Korean red ginseng) by HPLC-ELSD and its application to quality control. J Pharm Biomed Anal 2007;45:164-170. https://doi.org/10.1016/j.jpba.2007.05.001
  23. Wan JB, Lai CM, Li SP, Lee MY, Kong LY, Wang YT. Simultaneous determination of nine saponins from Panax notoginseng using HPLC and pressurized liquid extraction. J Pharm Biomed Anal 2006;41:274-279. https://doi.org/10.1016/j.jpba.2005.10.023
  24. Wan JB, Li P, Li S, Wang Y, Dong TT, Tsim KW. Simultaneous determination of 11 saponins in Panax notoginseng using HPLC-ELSD and pressurized liquid extraction. J Sep Sci 2006;29:2190-2196. https://doi.org/10.1002/jssc.200600103
  25. Cao H, Yu R, Choi Y, Ma ZZ, Zhang H, Xiang W, Lee DY, Berman BM, Moudgil KD, Fong HH et al. Discovery of cyclooxygenase inhibitors from medicinal plants used to treat inflammation. Pharmacol Res 2010;61:519-524. https://doi.org/10.1016/j.phrs.2010.02.007
  26. Mackman N. Triggers, targets and treatments for thrombosis. Nature 2008;451:914-918. https://doi.org/10.1038/nature06797
  27. Gambaryan S, Kobsar A, Rukoyatkina N, Herterich S, Geiger J, Smolenski A, Lohmann SM, Walter U. Thrombin and collagen induce a feedback inhibitory signaling pathway in platelets involving dissociation of the catalytic subunit of protein kinase A from an NFkappaB-IkappaB complex. J Biol Chem 2010;285:18352-18363. https://doi.org/10.1074/jbc.M109.077602
  28. Wang J, Huang ZG, Cao H, Wang YT, Hui P, Hoo C, Li SP. Screening of anti-platelet aggregation agents from Panax notoginseng using human platelet extraction and HPLC-DAD-ESI-MS/MS. J Sep Sci 2008;31:1173-1180. https://doi.org/10.1002/jssc.200700507
  29. Charo IF, Feinman RD, Detwiler TC. Interrelations of platelet aggregation and secretion. J Clin Invest 1977;60:866-873. https://doi.org/10.1172/JCI108841
  30. Park KM, Rhee MH, Shin HJ, Song YB, Hyun HC, Park KH, Cho HJ, Choi SA, Kang HC, Kim KJ et al. Inhibitory effects of panaxtriol from Panax ginseng C. A. Meyer on phosphoinositide breakdown induced by thrombin in platelets. J Ginseng Res 2008;32:107-113. https://doi.org/10.5142/JGR.2008.32.2.107
  31. Kimura Y, Okuda H, Arichi S. Effects of various ginseng saponins on 5-hydroxytryptamine release and aggregation in human platelets. J Pharm Pharmacol 1988;40:838-843. https://doi.org/10.1111/j.2042-7158.1988.tb06285.x
  32. Kuo SC, Teng CM, Lee JC, Ko FN, Chen SC, Wu TS. Antiplatelet components in Panax ginseng. Planta Med 1990;56:164-167. https://doi.org/10.1055/s-2006-960916
  33. Lee SR, Park JH, Choi KJ, Kim ND. Inhibitory effects of ginsenoside Rg3 on platelet aggregation and its mechanism of action. Korean J Ginseng Sci 1997;21:132-140.
  34. Eigenthaler M, Nolte C, Halbrugge M, Walter U. Concentration and regulation of cyclic nucleotides, cyclic-nucleotide-dependent protein kinases and one of their major substrates in human platelets. Estimating the rate of cAMP-regulated and cGMP-regulated protein phosphorylation in intact cells. Eur J Biochem 1992;205:471-481. https://doi.org/10.1111/j.1432-1033.1992.tb16803.x
  35. Ok WJ, Cho HJ, Kim HH, Lee DH, Kang HY, Kwon HW, Rhee MH, Kim M, Park HJ. Epigallocatechin-3-gallate has an anti-platelet effect in a cyclic AMP-dependent manner. J Atheroscler Thromb 2012;19:337-348. https://doi.org/10.5551/jat.10363
  36. Schwarz UR, Walter U, Eigenthaler M. Taming platelets with cyclic nucleotides. Biochem Pharmacol 2001; 62:1153-1161. https://doi.org/10.1016/S0006-2952(01)00760-2
  37. Kim S, Shim S, Choi DS, Kim JH, Kwon YB, Kwon J. Modulation of LPS-stimulated astroglial activation by ginseng total saponins. J Ginseng Res 2011;35:80-85. https://doi.org/10.5142/jgr.2011.35.1.080
  38. Shen T, Lee J, Park MH, Lee YG, Rho HS, Kwak YS, Rhee MH, Park YC, Cho JY. Ginsenoside Rp1, a ginsenoside derivative, blocks promoter activation of iNOS and COX-2 genes by suppression of an IKKb-mediated NF-kB pathway in HEK293 cells. J Ginseng Res 2011;35:200-208. https://doi.org/10.5142/jgr.2011.35.2.200
  39. Yayeh T, Jung KH, Jeong HY, Park JH, Song YB, Kwak YS, Kang HS, Cho JY, Oh JW, Kim SK et al. Korean red ginseng saponin fraction downregulates proinflammatory mediators in LPS stimulated RAW264.7 cells and protects mice against endotoxic shock. J Ginseng Res 2012;36:263-269. https://doi.org/10.5142/jgr.2012.36.3.263
  40. Lee JH, Park HJ. Effects of intaking of red ginseng products on human platelet aggregation and blood lipids. J Ginseng Res 1998;22:173-180.
  41. Jin YR, Yu JY, Lee JJ, You SH, Chung JH, Noh JY, Im JH, Han XH, Kim TJ, Shin KS et al. Antithrombotic and antiplatelet activities of Korean red ginseng extract. Basic Clin Pharmacol Toxicol 2007;100:170-175. https://doi.org/10.1111/j.1742-7843.2006.00033.x
  42. Yuan CS, Wang CZ, Wicks SM, Qi LW. Chemical and pharmacological studies of saponins with a focus on American ginseng. J Ginseng Res 2010;34:160-167. https://doi.org/10.5142/jgr.2010.34.3.160

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