Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Korean Red Ginseng protects endothelial cells from serum-deprived apoptosis by regulating Bcl-2 family protein dynamics and caspase S-nitrosylation

  • Kim, Young-Mi (Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University) ;
  • Kim, Jung Hwan (Department of Rehabilitation Medicine, Kangwon National University Hospital, School of Medicine, Kangwon National University) ;
  • Kwon, Hyuk Min (Department of Obstetrics and Gynecology, Kangwon National University Hospital) ;
  • Lee, Dong Heon (Department of Obstetrics and Gynecology, Kangwon National University Hospital) ;
  • Won, Moo-Ho (Department of Neurobiology, School of Medicine, Kangwon National University) ;
  • Kwon, Young-Guen (Department of Biochemistry, College of Sciences, Yonsei University) ;
  • Kim, Young-Myeong (Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University)
  • Received : 2013.05.06
  • Accepted : 2013.06.17
  • Published : 2013.10.15
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Abstract

Korean Red Ginseng extract (KRGE) is a traditional herbal medicine utilized to prevent endothelium dysfunction in the cardiovascular system; however, its underlying mechanism has not been clearly elucidated. We here examined the pharmacological effect and molecular mechanism of KRGE on apoptosis of human umbilical vein endothelial cells (HUVECs) in a serum-deprived apoptosis model. KRGE protected HUVECs from serum-deprived apoptosis by inhibiting mitochondrial cytochrome c release and caspase-9/-3 activation. This protective effect was significantly higher than that of American ginseng extract. KRGE treatment increased antiapoptotic Bcl-2 and Bcl-$X_L$ protein expression and Akt-dependent Bad phosphorylation. Moreover, KRGE prevented serum deprivation-induced subcellular redistribution of these proteins between the mitochondrion and the cytosol, resulting in suppression of mitochondrial cytochrome c release. In addition, KRGE increased nitric oxide (NO) production via Akt-dependent activation of endothelial NO synthase (eNOS), as well as inhibited caspase-9/-3 activities. These increases were reversed by co-treatment of cells with inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) and pre-incubation of cell lysates in dithiothreitol, indicating KRGE induces NO-mediated caspase modification. Indeed, KRGE inhibited caspase-3 activity via S-nitrosylation. These findings suggest that KRGE prevents serum deprivation-induced HUVEC apoptosis via increased Bcl-2 and Bcl-$X_L$ protein expression, PI3K/Akt-dependent Bad phosphorylation, and eNOS/NO-mediated S-nitrosylation of caspases. The cytoprotective property of KRGE may be valuable for developing new pharmaceutical means that limit endothelial cell death induced during the pathogenesis of vascular diseases.

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References

  1. Cines DB, Pollak ES, Buck CA, Loscalzo J, Zimmerman GA, McEver RP, Pober JS, Wick TM, Konkle BA, Schwartz BS et al. Endothelial cells in physiology and in the pathophysiology of vascular disorders. Blood 1998;91:3527-3561.
  2. Stoneman VE, Bennett MR. Role of apoptosis in atherosclerosis and its therapeutic implications. Clin Sci (Lond) 2004;107:343-354. https://doi.org/10.1042/CS20040086
  3. Williams TA, Verhovez A, Milan A, Veglio F, Mulatero P. Protective effect of spironolactone on endothelial cell apoptosis. Endocrinology 2006;147:2496-2505. https://doi.org/10.1210/en.2005-1318
  4. Desagher S, Martinou JC. Mitochondria as the central control point of apoptosis. Trends Cell Biol 2000;10:369-377. https://doi.org/10.1016/S0962-8924(00)01803-1
  5. Fujio Y, Nguyen T, Wencker D, Kitsis RN, Walsh K. Akt promotes survival of cardiomyocytes in vitro and protects against ischemia-reperfusion injury in mouse heart. Circulation 2000;101:660-667. https://doi.org/10.1161/01.CIR.101.6.660
  6. Stefanec T. Endothelial apoptosis: the missing link between atherosclerosis and SLE? Blood 2004;103:3608-3609. https://doi.org/10.1182/blood-2004-02-0714
  7. Kim YM, Bombeck CA, Billiar TR. Nitric oxide as a bifunctional regulator of apoptosis. Circ Res 1999;84:253-256. https://doi.org/10.1161/01.RES.84.3.253
  8. Lee SJ, Kim KM, Namkoong S, Kim CK, Kang YC, Lee H, Ha KS, Han JA, Chung HT, Kwon YG et al. Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53- dependent Noxa expression through the formation of Snitrosohomocysteine. J Biol Chem 2005;280:5781-5788. https://doi.org/10.1074/jbc.M411224200
  9. Hoffmann J, Haendeler J, Aicher A, Rossig L, Vasa M, Zeiher AM, Dimmeler S. Aging enhances the sensitivity of endothelial cells toward apoptotic stimuli: important role of nitric oxide. Circ Res 2001;89:709-715. https://doi.org/10.1161/hh2001.097796
  10. Sung J, Han KH, Zo JH, Park HJ, Kim CH, Oh BH. Effects of red ginseng upon vascular endothelial function in patients with essential hypertension. Am J Chin Med 2000;28:205-216. https://doi.org/10.1142/S0192415X00000258
  11. Kim YM, Namkoong S, Yun YG, Hong HD, Lee YC, Ha KS, Lee H, Kwon HJ, Kwon YG, Kim YM. Water extract of Korean red ginseng stimulates angiogenesis by activating the PI3K/Akt-dependent ERK1/2 and eNOS pathways in human umbilical vein endothelial cells. Biol Pharm Bull 2007;30:1674-1679. https://doi.org/10.1248/bpb.30.1674
  12. Lee SJ, Namkoong S, Kim YM, Kim CK, Lee H, Ha KS, Chung HT, Kwon YG, Kim YM. Fractalkine stimulates angiogenesis by activating the Raf-1/MEK/ERK- and PI3K/Akt/eNOS-dependent signal pathways. Am J Physiol Heart Circ Physiol 2006;291:H2836-H2846. https://doi.org/10.1152/ajpheart.00113.2006
  13. Qian J, Chen F, Kovalenkov Y, Pandey D, Moseley MA, Foster MW, Black SM, Venema RC, Stepp DW, Fulton DJ. Nitric oxide reduces NADPH oxidase 5 (Nox5) activity by reversible S-nitrosylation. Free Radic Biol Med 2012;52:1806-1819. https://doi.org/10.1016/j.freeradbiomed.2012.02.029
  14. Crow MT, Mani K, Nam YJ, Kitsis RN. The mitochondrial death pathway and cardiac myocyte apoptosis. Circ Res 2004;95:957-970. https://doi.org/10.1161/01.RES.0000148632.35500.d9
  15. Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis. Genes Dev 1999;13:1899-1911. https://doi.org/10.1101/gad.13.15.1899
  16. 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. https://doi.org/10.1038/21224
  17. Lu JM, Yao Q, Chen C. Ginseng compounds: an update on their molecular mechanisms and medical applications. Curr Vasc Pharmacol 2009;7:293-302. https://doi.org/10.2174/157016109788340767
  18. Yan J, Liu Q, Dou Y, Hsieh Y, Liu Y, Tao R, Zhu D, Lou Y. Activating glucocorticoid receptor-ERK signaling pathway contributes to ginsenoside Rg1 protection against $\beta$-amyloid peptide-induced human endothelial cells apoptosis. J Ethnopharmacol 2013;147:456-466. https://doi.org/10.1016/j.jep.2013.03.039
  19. Kwok HH, Ng WY, Yang MS, Mak NK, Wong RN, Yue PY. The ginsenoside protopanaxatriol protects endothelial cells from hydrogen peroxide-induced cell injury and cell death by modulating intracellular redox status. Free Radic Biol Med 2010;48:437-445. https://doi.org/10.1016/j.freeradbiomed.2009.11.013
  20. Min JK, Kim JH, Cho YL, Maeng YS, Lee SJ, Pyun BJ, Kim YM, Park JH, Kwon YG. 20(S)-ginsenoside Rg3 prevents endothelial cell apoptosis via inhibition of a mitochondrial caspase pathway. Biochem Biophys Res Commun 2006;349:987-994. https://doi.org/10.1016/j.bbrc.2006.08.129
  21. Dimmeler S, Haendeler J, Nehls M, Zeiher AM. Suppression of apoptosis by nitric oxide via inhibition of interleukin-1beta-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases. J Exp Med 1997;185:601-607. https://doi.org/10.1084/jem.185.4.601
  22. Sengupta S, Toh SA, Sellers LA, Skepper JN, Koolwijk P, Leung HW, Yeung HW, Wong RN, Sasisekharan R, Fan TP. Modulating angiogenesis: the yin and the yang in ginseng. Circulation 2004;110:1219-1225. https://doi.org/10.1161/01.CIR.0000140676.88412.CF
  23. Kim WY, Kim JM, Han SB, Lee SK, Kim ND, Park MK, Kim CK, Park JH. Steaming of ginseng at high temperature enhances biological activity. J Nat Prod 2000;63: 1702-1704. https://doi.org/10.1021/np990152b
  24. 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
  25. Zinkel S, Gross A, Yang E. BCL2 family in DNA damage and cell cycle control. Cell Death Differ 2006;13:1351-1359. https://doi.org/10.1038/sj.cdd.4401987
  26. Li H, Zhu H, Xu CJ, Yuan J. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 1998;94:491-501. https://doi.org/10.1016/S0092-8674(00)81590-1
  27. Zha J, Harada H, Yang E, Jockel J, Korsmeyer SJ. Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell 1996;87:619-628. https://doi.org/10.1016/S0092-8674(00)81382-3
  28. Moncada S, Radomski MW, Palmer RM. Endothelium-derived relaxing factor. Identification as nitric oxide and role in the control of vascular tone and platelet function. Biochem Pharmacol 1988;37:2495-2501. https://doi.org/10.1016/0006-2952(88)90236-5
  29. Shin W, Yoon J, Oh GT, Ryoo S. Korean red ginseng inhibits arginase and contributes to endotheliumdependent vasorelaxation through endothelial nitric oxide synthase coupling. J Ginseng Res 2013;37:64-73. https://doi.org/10.5142/jgr.2013.37.64
  30. Hellermann GR, Solomonson LP. Calmodulin promotes dimerization of the oxygenase domain of human endothelial nitric-oxide synthase. J Biol Chem 1997;272:12030-12034. https://doi.org/10.1074/jbc.272.18.12030
  31. Kwon YG, Min JK, Kim KM, Lee DJ, Billiar TR, Kim YM. Sphingosine 1-phosphate protects human umbilical vein endothelial cells from serum-deprived apoptosis by nitric oxide production. J Biol Chem 2001;276:10627-10633. https://doi.org/10.1074/jbc.M011449200
  32. Ceneviva GD, Tzeng E, Hoyt DG, Yee E, Gallagher A, Engelhardt JF, Kim YM, Billiar TR, Watkins SA, Pitt BR. Nitric oxide inhibits lipopolysaccharide-induced apoptosis in pulmonary artery endothelial cells. Am J Physiol 1998;275:L717-L728.
  33. Granger DL, Taintor RR, Cook JL, Hibbs JB Jr. Injury of neoplastic cells by murine macrophages leads to inhibition of mitochondrial respiration. J Clin Invest 1980;65:357-370. https://doi.org/10.1172/JCI109679
  34. Kim YM, Talanian RV, Billiar TR. Nitric oxide inhibits apoptosis by preventing increases in caspase-3-like activity via two distinct mechanisms. J Biol Chem 1997;272: 31138-31148. https://doi.org/10.1074/jbc.272.49.31138
  35. Mannick JB, Hausladen A, Liu L, Hess DT, Zeng M, Miao QX, Kane LS, Gow AJ, Stamler JS. Fas-induced caspase denitrosylation. Science 1999;284:651-654.s https://doi.org/10.1126/science.284.5414.651

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