Journal of Applied Biological Chemistry

  • 제65권1호
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  • Pages.57-62
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  • 2022
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  • 1976-0442(pISSN)
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  • 2234-7941(eISSN)
한국응용생명화학회 (The Korean Society for Applied Biological Chemistry)
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PI3K/Akt 및 MAPK 기전 조절을 통한 Artesunate의 콜라겐 유도의 사람 혈소판 응집 억제효과

Artesunate inhibits collagen-induced human platelets aggregation through regulation of PI3K/Akt and MAPK pathway

  • Lee, Dong-Ha (Department of Biomedical Laboratory Science, Molecular Diagnostics Research Institute, Namseoul University)
  • 투고 : 2022.01.17
  • 심사 : 2022.03.02
  • 발행 : 2022.03.31
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초록

혈소판의 과도한 활성화 및 응집은 심혈관계 질환의 주요원인이 된다. 따라서, 혈소판 활성화 및 응집을 억제하는 것은 심혈관계 질환을 예방하고 치료하는데 있어서 매력적인 치료 표적으로 여겨진다. 특히, 혈관 내피세포에서 분비되는 collagen에 의한 강력한 혈소판 활성화와 응집이 혈관질환에서 특징적이다. Artesunate는 Artemisia 또는 Scopolia 속 식물의 뿌리에서 추출한 화합물이며, 항암 및 알츠하이머 병 분야에서 효과가 있다고 보고된 바 있다. 그러나, artesunate가 collagen 유도의 혈소판 활성화와 응집에 미치는 영향과 그 기전에 대해서는 규명된 바가 없다. 본 연구에서는 collagen이 유도하는 사람 혈소판 응집에 있어 artesunate이 미치는 영향을 확인하였고, artesunate의 작용 기전을 명확히 하였다. Artesunate은 혈소판가 활성화 될 때 신호 전달 과정에 작용한다고 알려진 인단백질인 PI3K/Akt 및 MAPK의 인산화를 억제하였다. 또한, artesunate는 TXA2 생성을 감소시켰고, ATP 및 serotonin 방출 등의 혈소판 내 과립 분비를 감소시켰다. 그 결과, artesunate는 혈관 내피세포에서 분비되는 강력한 응집 유도 물질인 collagen으로 유도된 혈소판 응집을 106.41 μM의 IC50로 강력하게 억제하였다. 이 결과들을 통해, artesunate가 혈관 손상을 통해 일어나는 사람 혈소판의 활성화 및 응집을 억제하는 데 유효한 항혈전 물질로 가치가 있음을 제안한다.

Excessive activation and aggregation of platelets is a major cause of cardiovascular disease. Therefore, inhibition of platelet activation and aggregation is considered an attractive therapeutic target in preventing and treating cardiovascular diseases. In particular, strong platelet activation and aggregation by collagen secreted from the vascular endothelium are characteristic of vascular diseases. Artesunate is a compound extracted from the plant roots of Artemisia or Scopolia species, and has been reported to be effective in anticancer and Alzheimer's disease fields. However, the effect and mechanism of artesunate on collagen-induced platelet activation and aggregation have not been elucidated. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artesunate was clarified. Artesunate inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artesunate decreased TXA2 production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artesunate strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC50 of 106.41 µM. These results suggest that artesunate has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.

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참고문헌

  1. Jackson SP (2011) Arterial thrombosis-insidious, unpredictable and deadly. Nat Med 17: 1423-1436. doi: 10.1038/nm.2515
  2. Schwartz SM, Heimark RL, Majesky MW (1990) Developmental mechanisms underlying pathology of arteries. Physiol Rev 70: 1177-1209. doi: 10.1152/physrev.1990.70.4.1177
  3. Wonerow P, Obergfell A, Wilde JI, Bobe R, Asazuma N, Brdicka T, Leo A, Schraven B, Horejsi V, Shattil SJ, Watson SP (2002) Differential role of glycolipid-enriched membrane domains in glycoprotein VI- and integrin-mediated phospholipase Cgamma2 regulation in platelets. Biochemical Journal 364: 755-765. doi: 10.1042/bj20020128
  4. Payrastre B, Missy K, Trumel C, Bodin S, Plantavid M, Chap H (2000) The integrin alpha IIb/beta 3 in human platelet signal transduction. Biochem Pharmacol 60: 1069-1074. doi: 10.1016/S0006-2952(00)00417-2
  5. Morello F, Perino A, Hirsch E (2009) Phosphoinositide 3-kinase signalling in the vascular system. Cardiovasc Res 82: 261-271 https://doi.org/10.1093/cvr/cvn325
  6. Jennings LK (2009) Role of platelets in atherothrombosis. Am J Cardiol 103: 4A-10A https://doi.org/10.1016/j.amjcard.2008.11.017
  7. Sabatine MS, Jang IK (2000) The use of glycoprotein IIb/IIIa inhibitors in patients with coronary artery disease. Am J Med 109: 224-237 https://doi.org/10.1016/S0002-9343(00)00474-5
  8. Cattanco M, Tenconi PM, Lecchi A, Mannucci PM (1991) In vitro effects of picotamide on human platelet aggregation, the release reaction and thromboxane B2 production. Thromb Res 62: 717-724. doi: 10.1016/0049-3848(91)90375-7
  9. Su CY, Shiao MS, Wang CT (1999) Differential effects of ganodermic acid S on the thromboxane A2-signaling pathways in human platelets. Biochem Pharmacol 58: 587-595. doi: 10.1016/s0006-2952(99)00136-7
  10. Adam F, Kauskot A, Nurden P, Sulpice E, Hoylaerts MF, Davis RJ (2010) Platelet JNK1 is involved in secretion and thrombus formation. Blood 115: 4083-4092. doi: 10.1182/blood-2009-07-233932
  11. Bugaud F, Nadal-Wollbold F, Levy-Toledano S, Rosa JP, Bryckaert M (1999) Regulation of c-jun-NH2 terminal kinase and extracellular-signal regulated kinase in human platelets. Blood 94: 3800-3805 https://doi.org/10.1182/blood.v94.11.3800.423k25_3800_3805
  12. Kramer RM, Roberts EF, Strifler BA, Johnstone EM (1995) Thrombin induces activation of p38 MAP kinase in human platelets. J Biol Chem 270: 27395-27398. doi: 10.1074/jbc.270.46.27395
  13. Nadal-Wollbold F, Pawlowski M, Levy-Toledano S, Berrou E, Rosa JP, Bryckaert M (2002) Platelet ERK2 activation by thrombin is dependent on calcium and conventional protein kinases C but not Raf-1 or B-Raf. FEBS Lett 531: 475-482. doi: 10.1016/s0014-5793(02)03587-1
  14. Patrono C (1994) Aspirin as an antiplatelet drug. N Engl J Med 330: 1287-1294. doi: 10.1056/NEJM199405053301808
  15. Flevaris P, Li Z, Zhang G, Zheng Y, Liu J, Du X (2009) Two distinct roles of mitogen-activated protein kinases in platelets and a novel Rac1-MAPK-dependent integrin outside-in retractile signaling pathway. Blood 113: 893-901. doi: 10.1182/blood-2008-05-155978
  16. Kramer RM, Roberts EF, Um SL, Borsch-Haubold AG, Watson SP, Fisher MJ (1996) p38 mitogen-activated protein kinase phosphorylates cytosolic phospholipase A2 (cPLA2) in thrombin-stimulated platelets. Evidence that proline-directed phosphorylation is not required for mobilization of arachidonic acid by cPLA2. J Biol Chem 271: 27723-27729. doi: 10.1074/jbc.271.44.27723
  17. McNicol A, Shibou TS (1998) Translocation and phosphorylation of cytosolic phospholipase A2 in activated platelets. Thromb Res 92: 19-26. doi: 10.1016/s0049-3848(98)00097-8
  18. Chuang WY, Kung PH, Kuo CY, Wu CC (2013) Sulforaphane prevents human platelet aggregation through inhibiting the phosphatidylinositol 3-kinase/Akt pathway. Thromb Haemost 109: 1120-1130. doi: 10.1160/TH12-09-0636
  19. Thanaketpaisarn O, Waiwut P, Sakurai H, Saiki I (2011) Artesunate enhances TRAIL-induced apoptosis in human cervical carcinoma cells through inhibition of the NF-κB and PI3K/Akt signaling pathways. Int J Oncol 39: 279-285. doi: 10.3892/ijo.2011.1017
  20. Lu H, Wang B, Cui N, Zhang Y (2018) Artesunate suppresses oxidative and inflammatory processes by activating Nrf2 and ROS-dependent p38 MAPK and protects against cerebral ischemia-reperfusion injury. Mol Med Rep 17: 6639-6646. doi: 10.3892/mmr.2018.8666
  21. Shin JH, Kwon HW, Lee DH (2019) Ginsenoside F4 inhibits platelet aggregation and thrombus formation by dephosphorylation of IP3RI and VASP. J Appl Biol Chem 62: 93-100 https://doi.org/10.3839/jabc.2019.014
  22. Chang MC, Wang TM, Yeung SY, Jeng PY, Liao CH, Lin CC, Lin BR, Jeng JH (2011) Antiplatelet effect by p-cresol, a uremic and environmental toxicant, is related to inhibition of reactive oxygen species, ERK/p38 signaling and thromboxane A2 production. Atherosclerosis 219: 559-565. doi: 10.1016/j.atherosclerosis.2011.09.031
  23. Cipollone F, Patrignani P, Greco A, Panara MR, Padovano R, Cuccurullo F, Patrono C, Rebuzzi AG, Liuzzo G, Quaranta G, Maseri A (1997) Differential suppression of thromboxane biosynthesis by indobufen and aspirin in patients with unstable angina. Circulation 96: 1109-1116 https://doi.org/10.1161/01.CIR.96.4.1109
  24. Patrono C (2001) Aspirin: new cardiovascular uses for an old drug. Am J Med 110: 62S-65S. doi: 10.1016/s0002-9343(00)00645-8
  25. Calderwood DA (2004) Integrin activation. J Cell Sci 117: 657-666 https://doi.org/10.1242/jcs.01014