Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
권호 표지
DOI QR코드

DOI QR Code

Cudrania Tricuspidata root extract (CTE) has an anti-platelet effect via cGMP-dependent VASP phosphorylation in human platelets

꾸지뽕나무 뿌리 추출물의 cGMP에 의한 VASP 인산화 기전을 통한 항혈소판 효과

  • Ro, Ju-Ye (Department of Biomedical Laboratory Science, Konyang University) ;
  • Cho, Hyun-Jeong (Department of Biomedical Laboratory Science, Konyang University)
  • 노주예 (건양대학교 의과학대학 임상병리학과) ;
  • 조현정 (건양대학교 의과학대학 임상병리학과)
  • Received : 2019.10.31
  • Accepted : 2019.12.06
  • Published : 2019.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Cudrania tricuspidata has been reported to have many biological activities, including anti-inflammatory, anti-cancer, and antioxidant properties. However, the effects of C. tricuspidata root extract (CTE) on human platelet aggregation induced by collagen as well as the signaling pathways involved remain unknown. In the present study, we investigated the effect of CTE on human platelets. CTE inhibited platelet aggregation via down-regulation of thromboxane A2 (TXA2) by blocking cyclooxygenase-1 (COX-1) activity and intracellular Ca2+ mobilization in collagen-induced platelets. CTE also reduced the phosphorylation of phospholipase C (PLC) γ2 and syk. CTE regulated platelet aggregation via cyclic guanosine monophosphate (cGMP)-dependent phosphorylation of vasodilator-stimulated phosphoprotein (VASP) Ser239. In addition, administration of CTE (50 and 100 mg/kg) significantly reduced hyper-aggregated platelet aggregation by collagen (5 ㎍/mL) without hepatotoxicity in HFD (high fat diet)-fed rats. Taken together, these results suggest that CTE has anti-platelet effects both in vitro and in vivo. Therefore, CTE may be an effective therapeutic and preventive agent for cardiovascular disease, and is a safe and natural product.

꾸지뽕나무 뿌리 추출물은 항염, 항암, 항산화와 같은 효과를 갖는 많은 생리활성 물질을 포함하고 있다고 알려져 있다. 그러나 꾸지뽕나무 뿌리 추출물(이하 CTE)의 사람 혈소판 응집 억제 기전에 관하여는 알려진 바 없다. 본 연구에서는 CTE가 혈소판에 미치는 영향을 확인하고자 하였다. CTE는 collagen으로 유도한 혈소판 응집 반응에서 cyclooxygenase-1 활성을 억제하고, 세포 내 칼슘 농도를 낮추는 방식으로 thromboxane A2 생성을 억제하였다. 또한, phospholipase Cγ2와 syk의 인산화 반응을 억제하였으며, guanosine monophosphate (cGMP)에 의존적인 방식으로 vasodilator-stimulated phosphoprotein(VASP)의 Ser239 위치를 인산화하여 항혈소판 효과를 나타내었다. 또한, 고지방 식이로 혈소판 활성화를 유도한 랫드에서 CTE를 경구 투여 하였을 때, 간독성 없이 콜라겐으로 유도한 혈소판 응집반응을 thromboxane A2 생성을 억제함으로써 항혈소판 효과를 보였다. 이는 in vivo와 in vitro에서 같은 결과를 제시하고 있다. 결론적으로, CTE는 항혈소판 작용 및 심혈관계 질환 예방을 위한 천연물 유래의 안전하고, 새로운 물질임을 제시하는 바이다.

Keywords

References

  1. M. Marin, S. Manez, "Recent trends in the pharmacological activity of isoprenyl phenolics", Curr Med Chem, Vol.20, No.2, pp.272-279, 2013. DOI: https://doi.org/10.2174/092986713804806676
  2. X. Li, Z. Yao, X. Jiang, J. Sun, G. Ran, X. Yang, Y. Zhao, Y. Yan, Z. Chen, L. Tian, W. Bai, "Bioactive compounds from Cudrania tricuspidata : A natural anticancer source", Crit Rev Food Sci Nutr, Vol.22, pp.1-21, Dec, 2018. DOI: https://doi.org/10.1080/10408398.2018.1541866
  3. E. J. Seo, M. J. Curtis-Long, B. W. Lee, H. Y. Kim, Y. B. Ryu, T. S. Jeong, W. S. Lee, K. H. Park, "Xanthones from Cudrania tricuspidata displaying potent alphaglucosidase inhibition", Bioorg Med Chem Lett, Vol.17, No.23, pp.6421-6424, Dec, 2007. DOI: https://doi.org/10.1016/j.bmcl.2007.10.00
  4. H. Y. Joo, K. T. Lim, "Glycoprotein isolated from Cudrania tricuspidata Bureau inhibits iNO and COX-2 expression through modulation of NF-kappaB in LPS-stimulated RAW 264.7 cells", Environ Toxicol Pharmacol, Vol.27, No.2, pp.247-252, Mar, 2009. DOI: https://doi.org/10.1016/j.etap.2008.10.014
  5. T. S. Jeong, Y. B. Ryu, H. Y. Kim, M. J. Curtis-Long, S. J. An, J. H. Lee, W. S. Lee, K. H. Park, "Low density lipoprotein (LDL)-antioxidant flavonoids from roots of Sophora flavescens", Biol Pharm Bull, Vol. 31, No. 11, pp. 2097-2102, Nov, 2008. DOI: https://doi.org/10.1248/bpb.31.2097
  6. H. Yoo, S. K. Ku, W. Lee, S. Kwak, Y. D. Baek, B. W. Min, G. S. Jeong and J. S. Bae, "Antiplatelet, anticoagulant, and profibrinolytic activities of cudratricus xanthone A", Arch Pharm Res, Vol.37, No.8, pp.1069-1078, Aug, 2014. DOI: https://doi.org/10.1007/s12272-013-0290-4
  7. S. Geue, B. Walker-Allgaier, D. Eissler, R. Tegt- meyer, M. Schaub, F. Lang, M. Gawaz, O, Borst, P. Munzer, "Doxepin inhibits GPVI-dependent platelet $Ca^{2+}$ signaling and collagen-dependent thrombus formation", Am J Physiol Cell Physiol, Vol.312, No.6, pp.C765-C774, Apr, 2017. DOI: https://doi.org/10.1152/ajpcell.00262.2016
  8. W. Bergmeier, L. Stefanini, "Platelet ITAM signaling", Curr Opin Hematol, Vol.20, No.5, pp.445-450, Sep, 2013. DOI: https://doi.org/10.1097/MOH.0b013e3283642267
  9. R. H. Lee, W. Bergmeier, "Platelet immunoreceptor tyrosine-based activation motif (ITAM) and hemITAM signaling and vascular integrity in inflammation and development", J Thromb Haemost, Vol.14, No.4, pp.645-654, Apr, 2016. DOI: https://doi.org/10.1111/jth
  10. C. Feng, C. B. Post, "Insights into the allosteric regulation of Syk association with receptor ITAM, a multi-state equilibrium", Phys Chem Chem Phys, Vol.18, No.8, pp.5807-5818, Feb, 2016. DOI: https://doi.org/10.1039/c5cp05417f
  11. J. H. Ryu, J. Y. Ro, H. J. Park, H. J. Cho, "Anti-platelet effect of Ginkgolide A from Ginkgo biloba ", J Korean Soc Appl Biol Chem, Vol.57, No.2, pp.221-228, Apr, 2014. DOI: http://doi.org/10.1007/s13765-013-4275-2
  12. H. J. Cho, H. J. Kang, Y. J. Kim, D. H. Lee, H. W. Kwon, Y. Y. Kim, H. J. Park, "Inhibition of platelet aggregation by chlorogenic acid via cAMP and cGMP-dependent manner", Blood Coagul Fibrinolysis, Vol.23, No.7, pp.629-635, Oct, 2012. DOI: https://doi.org/10.1097/MBC.0b013e3283570846
  13. J. H. Shin, H. W. Kwon, H. J. Cho, M. H. Rhee, H. J. Park, "Vasodilator-stimulated phosphoprotein-phosphorylation by ginsenoside Ro inhibits fibrinogen binding to aIIb/b3 in thrombin-induced human platelets", J Ginseng Res, Vol.40, pp.359-365, 2016. DOI: http://doi.org/10.1016/j.jgr.2015.11.003
  14. Y. Wu, S. J. Gunst, "Vasodilator-stimulated phosphoprotein (VASP) regulates actin polymerization and contraction in airway smooth muscle by a vinculindependent mechanism", J Biol Chem, Vol.290, No.18, pp.11403-11416, May, 2015. DOI: https://doi.org/10.1074/jbc.M115.645788
  15. J. Y. Ro, J. H. Ryu, H. J. Park, H. J. Cho, "Onion (Allium cepa L.) peel extract has anti-platelet effects in rat platelets", SpringerPlus, Vol.4, No.7, pp.1-8, Jan, 2015. DOI: https://doi.org/10.1186/s40064-015-0786-0
  16. D. S. Bush, R. L. Jones, "Measurement of cytoplasmic calcium in aleurone protoplasts using indo-1 and fura-2; Progress and Prospects", Cell Calcium, Vol.93, No.3, pp.841-845, Jul, 1990. DOI: https://doi.org/10.1104/pp.93.3.841
  17. K. Anders, "Resolution of Students t-tests, ANOVA and analysis of variance components from intermediary data", Biochem Med (Zagreb), Vol.27, No.2, pp.253-258. Jun, 2017. DOI: https://doi.org/10.11613/BM.2017.026
  18. J. H. Shin, H. W. Kwon, M. H. Rhee, H. J. Park, "Inhibitory effects of thromboxane $A_{2}$ generation by ginsenoside Ro due to attenuation of cytosolic phospholipase $A_{2}$ phosphorylation and arachidonic acid release", J Ginseng Res, Vol.43, No.2, pp.236-241, Apr, 2019. DOI: https://doi.org/10.1016/j.jgr.2017.12.007
  19. D. H. Lee, Y. J. Kim, H. H. Kim, H. J. Cho, J. H. Ryu, M. H. Rhee, H. J. Park, "Inhibitory effects of epigallocatechin-3-gallate on microsomal cyclooxygenase-1 activity in platelets", Vol.21, no.1, pp.54-59, Jan, 2013. DOI: https://doi.org/10.4062/biomolther.2012.075
  20. R. Lorenz, D. Betinetti, F. W. Herberg, "cAMPDependent Protein Kinase and cGMP-Dependent Protein Kinase as Cyclic Nucleotide Effectors", Handb Exp Pharmacol, Vol.238, pp.105-122, 2017. DOI: https://doi.org/10.1007/164_2015_36
  21. H. Jahn, W. Nastainczyk, A. Rohrkasten, T. Schneider, F. Hofmann, "High-Throughput $Ca^{2+}$ Flux Assay To Monitor CyclicNucleotide-Gated Channel Activity and Characterize Achromatopsia Mutant Channel Function", ACS Chem Neurosci, Vol.10, No.8, pp.3662-3670, Aug, 2019. DOI: https://doi.org/10.1021/acschemneuro.9b00231
  22. P. F. Monteiro, R. P. Morganti, M. A. Delbin, M. C. Calixto, M. E. Lopes-Pires, S. Marcondes, A. Zanesco, E. Antunes, "Platelet hyperaggregability in high-fat fed rats: a role for intraplatelet reactive-oxygen species production", Cardiovasc Diabetol, Vol.11, pp.5, Jan, 2012. DOI: https://doi.org/10.1186/1475-2840-11-5