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Comparative anti-thrombotic activity and haemorrhagic adverse effect of nattokinase and tissue-type plasminogen activator

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Abstract

Anti-thrombotic activity and safety of nattokinase, an enzyme produced by Bacillus subtilis during soybean fermentation, were investigated in comparison with tissue-type plasminogen activator (t-PA). Carotid arterial thrombosis was produced with a FeCl3-soaked paper, followed by intravenous injection of nattokinase or t-PA. Nattokinase and t-PA delayed thrombus formation, near-fully (> 90%) inhibiting at 75 and 8.5 mg/kg, respectively. As adverse effects, t-PA induced petechial haemorrhage at 10 mg/kg in the lungs and thymus, and extensive bleeding at 20 mg/kg. Nattokinase also caused pulmonary haemorrhage from 300 mg/kg. Collectively, the standard safety margins (SSMs) for t-PA and nattokinase were calculated to be 1.2 and 4.0, respectively. Combinational treatment with dexamethasone (2 mg/kg) increased the efficacy and safety of t-PA and nattokinase, widening their SSMs to 2.4 and 8.0, respectively. The results indicate that nattokinase delayed thrombus formation and dissolved thrombi, and that nattokinase could be a good candidate anti-thrombotic agent with relatively-low haemorrhagic risk.

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References

  • Brass LM, Lichtman JH, Wang Y, Gurwitz JH, Radford MJ, Krumholz HM. Intracranial hemorrhage associated with thrombolytic therapy for elderly patients with acute myocardial infarction. Stroke 31: 1802–1811 (2000)

    Article  CAS  Google Scholar 

  • Bronicki RA, Backer CL, Baden HP, Mavroudis C, Crawford SE, Green TP. Dexamethasone reduces the inflammatory response to cardiopulmonary bypass in children. Ann. Thorac. Surg. 69: 1490–1495 (2000)

    Article  CAS  Google Scholar 

  • Del Zoppo GJ, Saver JL, Jauch EC, Adams HP Jr; American Heart Association Stroke Council. Expansion of the time window for treatment of acute ischemic stroke with intravenous tissue plasminogen activator: a science advisory from the American Heart Association/American Stroke Association. Stroke 40: 2945–2948 (2009)

    Article  Google Scholar 

  • Eckly A, Hechler B, Freund M, Zerr M, Cazenave JP, Lanza F, Mangin PH, Gachet C. Mechanisms underlying FeCl3-induced arterial thrombosis. J. Thromb. Haemost. 9: 779–789 (2011)

    Article  CAS  Google Scholar 

  • El Azab SR, Rosseel PM, de Lange JJ, Groeneveld AB, van Strik R, van Wijk EM, Scheffer GJ. Dexamethasone decreases the pro- to anti-inflammatory cytokine ratio during cardiac surgery. Br. J. Anaesth. 88: 496–501 (2002)

    Article  Google Scholar 

  • Emberson J, Lees KR, Lyden P, Blackwell L, Albers G, Bluhmki E, Brott T, Cohen G, Davis S, Donnan G, Grotta J, Howard G, Kaste M, Koga M, von Kummer R, Lansberg M, Lindley RI, Murray G, Olivot JM, Parsons M, Tilley B, Toni D, Toyoda K, Wahlgren N, Wardlaw J, Whiteley W, del Zoppo GJ, Baigent C, Sandercock P, Hacke W; Stroke Thrombolysis Trialists’ Collaborative Group. Effect of treatment delay, age, and stroke severity on the effects of intravenous thrombolysis with alteplase for acute ischaemic stroke: a meta-analysis of individual patient data from randomised trials. Lancet 384: 1929–1935 (2014)

    Article  CAS  Google Scholar 

  • Fischer S, Renz D, Schaper W, Karliczek GF. In vitro effects of dexamethasone on hypoxia-induced hyperpermeability and expression of vascular endothelial growth factor. Eur. J. Pharmacol. 411: 231–243 (2001)

    Article  CAS  Google Scholar 

  • Fujita M, Hong K, Ito Y, Fujii R, Kariya K, Nishimuro S. Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat. Biol. Pharm. Bull. 18: 1387–1391 (1995)

    Article  CAS  Google Scholar 

  • Fujita M, Ohnishi K, Takaoka S, Ogasawara K, Fukuyama R, Nakamuta H. Antihypertensive effects of continuous oral administration of nattokinase and its fragments in spontaneously hypertensive rats. Biol. Pharm. Bull. 34: 1696–1701 (2011)

    Article  CAS  Google Scholar 

  • Furie B, Furie BC. Thrombus formation in vivo. J. Clin. Invest. 115: 3355–3362 (2005)

    Article  CAS  Google Scholar 

  • Heying R, Wehage E, Schumacher K, Tassani P, Haas F, Lange R, Hess J, Seghaye MC. Dexamethasone pretreatment provides antiinflammatory and myocardial protection in neonatal arterial switch operation. Ann. Thorac. Surg. 93: 869–876 (2012)

    Article  Google Scholar 

  • Hitosugi M, Hamada K, Misaka K. Effects of Bacillus subtilis var. natto products on symptoms caused by blood flow disturbance in female patients with lifestyle diseases. Int. J. Gen. Med. 8: 41–66 (2015)

    Article  CAS  Google Scholar 

  • Jackson SP, Nesbitt WS, Kulkarni S. Signaling events underlying thrombus formation. J. Thromb. Haemost. 1: 1602–1612 (2003)

    Article  CAS  Google Scholar 

  • Jang JY, Kim TS, Cai J, Kim J, Kim Y, Shin K, Kim KS, Park SK, Lee SP, Choi EK, Rhee MH, Kim YB. Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. Lab. Anim. Res. 29: 221–225 (2013)

    Article  Google Scholar 

  • Jang JY, Kim TS, Cai J, Kim J, Kim Y, Shin K, Kim KS, Lee SP, Kang MH, Choi EK, Rhee MH, Kim YB. Perilla oil improves blood flow through inhibition of platelet aggregation and thrombus formation. Lab. Anim. Res. 30: 21–27 (2014)

    Article  Google Scholar 

  • Jin X, Liu J, Liu W. Early ischemic blood brain barrier damage: a potential indicator for hemorrhagic transformation following tissue plasminogen activator (tPA) thrombolysis? Curr. Neurovasc. Res. 11: 254–262 (2014)

    Article  CAS  Google Scholar 

  • Kamiya S, Hagimori M, Ogasawara M, Arakawa M. In vivo evaluation method of the effect of nattokinase on carrageenan-induced tail thrombosis in a rat model. Acta Haematol. 124: 218–224 (2010)

    Article  CAS  Google Scholar 

  • Kim JY, Gum SN, Paik JK, Lim HH, Kim KC, Ogasawara K, Inoue K, Park S, Jang Y, Lee JH. Effects of nattokinase on blood pressure: a randomized, controlled trial. Hypertens. Res. 31: 1583–1588 (2008)

    Article  CAS  Google Scholar 

  • Ko JH, Yan JP, Zhu L, Qi YP. Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK02. Comp. Biochem. Physiol. C Toxicol. Pharmacol. 137: 65–74 (2004)

    Article  Google Scholar 

  • Konstantinides S, Schafer K, Thinnes T, Loskutoff DJ. Plasminogen activator inhibitor-1 and its cofactor vitronectin stabilize arterial thrombi after vascular injury in mice. Circulation 103: 576–583 (2001)

    Article  CAS  Google Scholar 

  • Kotb E. The biotechnological potential of fibrinolytic enzymes in the dissolution of endogenous blood thrombi. Biotechnol. Prog. 30: 656–672 (2014)

    Article  CAS  Google Scholar 

  • Lee JJ, Yang H, Yoo YM, Hong SS, Lee D, Lee HJ, Lee HJ, Myung CS, Choi KC, Jeung EB. Morusinol extracted from Morus alba inhibits arterial thrombosis and modulates platelet activation for the treatment of cardiovascular disease. J. Atheroscler. Thromb. 19: 516–522 (2012)

    Article  CAS  Google Scholar 

  • Majid A, Delanty N, Kantor J. Antiplatelet agents for secondary prevention of ischemic stroke. Ann. Pharmacother. 35: 1241–1247 (2001)

    Article  CAS  Google Scholar 

  • Nieto Estrada VH, Molano Franco D, Medina RD, Gonzalez Garay AG, Martí-Carvajal AJ, Arevalo-Rodriguez I. Interventions for preventing high altitude illness: Part 1. Commonly-used classes of drugs. Cochrane Database Syst. Rev. 6: CD009761 (2017)

    PubMed  Google Scholar 

  • Rudolf J, Grond M, Prince WS, Schmülling S, Heiss WD. Evidence of anaphylaxy after alteplase infusion. Stroke 30: 1142–1143 (1999)

    Article  CAS  Google Scholar 

  • Sumi H, Hamada H, Nakanishi K, Hiratani H. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematol. 84: 139–143 (1990)

    Article  CAS  Google Scholar 

  • Sumi H, Hamada H, Tsushima H, Mihara H, Muraki H. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet. Experientia 43: 1110–1111 (1987)

    Article  CAS  Google Scholar 

  • Suzuki Y, Kondo K, Matsumoto Y, Zhao BQ, Otsuguro K, Maeda T, Tsukamoto Y, Urano T, Umemura K. Dietary supplementation of fermented soybean, natto, suppresses intimal thickening and modulates the lysis of mural thrombi after endothelial injury in rat femoral artery. Life Sci 73: 1289–1298 (2003)

    Article  CAS  Google Scholar 

  • The National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group. Tissue plasminogen activator for acute ischemic stroke. N. Engl. J. Med. 333: 1581–1587 (1995)

    Article  Google Scholar 

  • Urano T, Ihara H, Umemura K, Suzuki Y, Oike M, Akita S, Tsukamoto Y, Suzuki I, Takada A. The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis Cleaves and inactivates plasminogen activator inhibitor type 1. J. Biol. Chem. 276: 24690–24696 (2001)

    Article  CAS  Google Scholar 

  • Xu J, Du M, Yang X, Chen Q, Chen H, Lin DH. Thrombolytic effects in vivo of nattokinase in a carrageenan-induced rat model of thrombosis. Acta Haematol. 132: 247–253 (2014)

    Article  CAS  Google Scholar 

  • Zafren K. Prevention of high altitude illness. Travel Med. Infect. Dis. 12: 29–39 (2014)

    Article  Google Scholar 

  • Zhang Y, Houng A, Reed G. Synergistic inhibitors halt brain and systemic hemorrhage after tissue plasminogen activator treatment for ischemic stroke. Stroke 44: ATP260 (2013)

    Article  Google Scholar 

  • Zivin JA. Thrombolytic stroke therapy: past, present, and future. Neurology 53: 14–19 (1999)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (2017R1A2A2A05069417).

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Authors and Affiliations

  1. College of Veterinary Medicine and Veterinary Medical Center, Chungbuk National University, 1 Chungdaero (Gaesin-dong), Cheongju, Chungbuk, 28644, Korea

    Haiyu Guo, Young-Hwan Ban, Yeseul Cha, Eun Suk An, Jieun Choi, Da Woom Seo, Ehn-Kyoung Choi & Yun-Bae Kim

  2. Department of Biology Education, Korea National University of Education, Cheongju, Chungbuk, 28173, Korea

    Dongsun Park

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  1. Haiyu Guo
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  2. Young-Hwan Ban
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Guo, H., Ban, YH., Cha, Y. et al. Comparative anti-thrombotic activity and haemorrhagic adverse effect of nattokinase and tissue-type plasminogen activator. Food Sci Biotechnol 28, 1535–1542 (2019). https://doi.org/10.1007/s10068-019-00580-1

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