Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Inhibitory effects of artemether on thrombus formation via regulation of cyclic nucleotides in collagen-induced platelets

콜라겐-유도의 혈소판에서 사이클릭 뉴클레오티드의 조절을 통한 Artemether의 항혈전 효과

  • Chang-Eun, Park (Department of Biomedical Laboratory Science, Molecular Diagnostics Research Institute, Namseoul University) ;
  • Dong-Ha, Lee (Department of Biomedical Laboratory Science, Molecular Diagnostics Research Institute, Namseoul University)
  • Received : 2022.08.26
  • Accepted : 2022.09.26
  • Published : 2022.12.31
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Abstract

Although normal activation of platelets is important in the process of hemostasis, excessive or abnormal activation of platelets can lead to cardiovascular diseases. Therefore, the discovery of novel substances capable of regulating or inhibiting platelet activation may be helpful in the prevention and treatment of cardiovascular diseases. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. This study investigated the effects of artemether on platelet activation and thrombus formation induced by collagen. As a result, cAMP level was significantly increased by artemether, and VASP and IP3R, substrates of cAMP-dependent kinase, were phosphorylated. IP3R phosphorylation by Artemether inhibited Ca2+ recruitment into the cytoplasm, and phosphorylated VASP inhibited fibrinogen binding by inactivating αIIb/β3 located on the platelet membrane. Consequently, artemether inhibited thrombin-induced fibrin clot formation. Therefore, we propose that artemether can act as an effective prophylactic and therapeutic agent for cardiovascular diseases caused by excessive platelet activation and thrombus formation.

지혈이 일어나는 과정에서 혈소판의 정상적인 활성화가 중요하지만, 혈소판의 과도하거나 비정상적인 활성화는 뇌졸중, 죽상동맥 경화증 및 혈전증과 같은 심혈관 질환을 유발할 수 있다. 따라서, 혈소판 활성화를 조절하거나 억제할 수 있는 새로운 물질의 발견은 심혈관 질환의 예방 및 치료에 도움이 될 수 있다. Artemether는 Artemisia annua의 유효성분으로 알려진 artemisinin의 유도체로서 말라리아 치료에 효과적이라고 보고된 바 있고, 항산화 및 대사 효소 억제를 통해 기능하는 것으로 알려져 있다. 그런데, 혈소판 활성화 및 응집에 있어서의 artemether의 역할과 collagen으로 유도한 사람 혈소판에서 artemether의 작용기전은 현재까지 알려진 바가 없다. 본 연구는 artemether가 collagen에 의해 유도된 혈소판 활성화와 혈전 형성에 어떤 영향을 미치는 지 연구하였다. 그 결과, cAMP level이 artemether에 의해 유의하게 증가되었고, cAMP-의존성 kinase의 기질인 VASP 및 IP3R이 인산화되었다. Artemether에 의한 IP3R 인산화는 세포질 내로의 Ca2+ 동원을 억제하였고, 인산화된 VASP는 혈소판 막에 위치한 αIIb/β3 불활성화를 일으켜 fibrinogen 결합을 억제하였다. 결과적으로, artemether는 thrombin으로 유도한 fibrin clot의 형성을 억제하였다. 따라서, 우리는 artemether가 과도한 혈소판 활성화 및 혈전 형성으로 인한 심혈관 질환의 효과적인 예방 및 치료제로 작용할 수 있음을 제안한다.

Keywords

References

  1. Thompson RC, Allam AH, Lombardi GP, Wann LS, Sutherland M., Sutherland JD, Soliman TT, Frohlich B, Mininberg DT, Monge JM, Vallodolid CM, Cox SL, Abd el-Maksoud G, Badr I, Miyamoto MI, el-Halim Nur el-Din A, Narula J, Finch CE, Thomas GS (2013) Atherosclerosis across 4000 years of human history: the Horus study of four ancient populations. Lancet 381: 1211-1222. doi: 10.1016/S0140-6736(13)60598-X 
  2. Rouzer CA, Marnett LJ (2005) Structural and functional differences between cyclooxygenases: fatty acid oxygenases with a critical role in cell signaling. Biochem Biophys Res Commun 338: 34-44. doi: 10.1016/j.bbrc.2005.07.198 
  3. Li Z, Delaney MK, O'Brien KA, Du X (2010) Signaling during platelet adhesion and activation. Arterioscler. Thromb. Vasc. Biol. 30: 2341-2349. doi: 10.1161/ATVBAHA.110.207522 
  4. Schwarz UR, Walter U, Eigenthaler M (2001) Taming platelets with cyclic nucleotides. Biochem Pharmacol 62: 1153-1161. doi: 10.1016/S0006-2952(01)00760-2 
  5. Quinton TM, Dean WL (1992) Cyclic AMP-dependent phosphorylation of the inositol-1,4,5-trisphosphate receptor inhibits Ca2+ release from platelet membranes. Biochemical and Biochem Biophys Res Commun 184: 893-899. doi: 10.1016/0006-291X(92)90675-B 
  6. Cavallini L, Coassin M, Borean A, Alexandre A (1996) Prostacyclin and sodium nitroprusside inhibit the activity of the platelet inositol 1,4,5-trisphosphate receptor and promote its phosphorylation. J Biol Chem 271: 5545-5551. doi: 10.1074/jbc.271.10.5545 
  7. Laurent V, Loisel TP, Harbeck B, Wehman A, Grobe L, Jockusch BM, Frank JW, Gertler B, Carlier MF (1999) Role of proteins of the Ena/VASP family in actin-based motility of Listeria monocytogenes. J Cell Biol 144: 1245-1258. doi: 10.1083/jcb.144.6.1245 
  8. Sudo T, Ito H, Kimura Y (2003) Phosphorylation of the vasodilator-stimulated phosphoprotein (VASP) by the anti-platelet drug, cilostazol, in platelets. Platelets 14: 381-390. doi: 10.1080/09537100310001598819 
  9. Ma YQ, Qin J, Plow EF (2007) Platelet integrin alpha(IIb)beta(3): activation mechanisms. J Thromb Haemost 5: 1345-1352. doi: 10.1111/j.1538-7836.2007.02537.x 
  10. 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 
  11. Esu EB, Effa EE, Opie ON, Meremikwu MM (2019) Artemether for severe malaria, Cochrane Database Syst Rev 6: CD010678. doi: 10.1002/14651858.CD010678.pub3 
  12. Saeed ME, Krishna S, Greten HJ, Kremsner PG, Efferth T (2016) Antischistosomal activity of artemisinin derivatives in vivo and in patients. Pharmacological Research 110: 216-226. doi: 10.1016/j.phrs.2016.02.017 
  13. Xu J, He X, Huang X, Zhang F, Ren X, Asakiya C, Li Y, Huang K (2022) Artemether Ameliorates Non-Alcoholic Steatohepatitis by Repressing Lipogenesis, Inflammation, and Fibrosis in Mice. Front Pharmacol 13: 851342. doi: 10.3389/fphar.2022.851342 
  14. Schaeffer J, Blaustein MP (1989) Platelet free calcium concentrations measured with fura-2 are influenced by the transmembrane sodium gradient. Cell Calcium 10: 101-113. doi: 10.1016/0143-4160(89)90050-X 
  15. Dong J, Yuan J, Wang JL, Ji RF, Quan QH, Guo XY, Gao J, Liu YG (2017) Study on screening antitumor active fractions and chemical components in active fractions from root of Anaycclus pyrethrum. Zhongguo Zhong Yao Za Zhi 42: 3932-3937. doi: 10.19540/j.cnki.cjcmm.2017.0158 
  16. 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. doi: 10.3839/jabc.2019.014 
  17. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260: 3440-3450. doi: 10.1016/S0021-9258(19)83641-4 
  18. Nishikawa M, Tanaka T, Hidaka H (1980) Ca2+-calmodulin-dependent phosphorylation and platelet secretion. Nature 287: 863-865. doi: 10.1038/287863a0 
  19. Kuo JF, Andersson RG, Wise BC, Mackerlova L, Salomonsson I, Brackett NL, Katoh N, Shoji M, Wrenn RW (1980) Calcium-dependent protein kinase: widespread occurrence in various tissues and phyla of the animal kingdom and comparison of effects of phospholipid, calmodulin, and trifluoperazine. Proc Natl Acad Sci 77: 7039-7043. doi: 10.1073/pnas.77.12.7039 
  20. Gambaryan S, Kobsar A, Rukoyatkina N, Herterich S, Geiger J, Smolenski A, Lohmann SM, Walter U (2010) 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 285: 18352-18363. doi: 10.1074/jbc.M109.077602 
  21. Wangorsch G, Butt E, Mark R, Hubertus K, Geiger J, Dandekar T, Dittrich M (2011) Time-resolved in silico modeling of finetuned cAMP signaling in platelets: feedback loops, titrated phospho-rylations and pharmacological modulation, BMC Syst Biol 5: 178. doi: 10.1186/1752-0509-5-178 
  22. Phillips DR, Nannizzi-Alaimo L, Prasad KS (2001) Beta3 tyrosine phosphorylation in alphaIIbbeta3 (platelet membrane GP IIb-IIIa) outside-in integrin signaling. Thromb Haemost 86: 246-258. doi: 10.1055/s-0037-1616222 
  23. Horstrup K, Jablonka B, Honig-Liedl P, Just M, Kochsiek K, Walter U (1994) Phosphorylation of focal adhesion vasodilator-stimulated phosphoprotein at Ser157 in intact human platelets correlates with fibrinogen receptor inhibition. Eur J Biochem 225: 21-27. doi: 10.1111/j.1432-1033.1994.00021.x 
  24. Singha P, Bajpai V, Khandelwal N, Varshney S, Gaikwadc AN, Srivastava M, Singhb B, Kumar B (2021) Determination of bioactive compounds of Artemisia Spp. plant extracts by LC-MS/MS technique and their in-vitro anti-adipogenic activity screening. J Pharm Biomed Anal 193:113707. doi: 10.1016/j.jpba.2020.113707