. 2015 Apr 21;10(4):e0123454.
doi: 10.1371/journal.pone.0123454. eCollection 2015.
Antonio Kokot 1 , Domagoj Drmic 2 , Masa Hrelec Patrlj 2 , Anita Zenko Sever 3 , Danijela Kolenc 3 , Bozo Radic 2 , Jelena Suran 2 , Davor Bojic 2 , Aleksandar Vcev 1 , Sven Seiwerth 3 , Predrag Sikiric 2
Affiliations
PMID: 25897838 PMCID: PMC4405609 DOI: 10.1371/journal.pone.0123454
Pentadecapeptide BPC 157 Reduces Bleeding and Thrombocytopenia after Amputation in Rats Treated with Heparin, Warfarin, L-NAME and L-Arginine
Mirjana Stupnisek et al. PLoS One. 2015.
Abstract
Background: BPC 157 is a stable gastric pentadecapeptide recently implicated with a role in hemostasis. While NO is largely implicated in hemostatic mechanisms, in tail-amputation-models under heparin- and warfarin-administration, both the NO-synthase (NOS)-blocker, L-NAME (prothrombotic) and the NOS-substrate L-arginine (antithrombotic), were little investigated. Objective. To investigate the effect of L-NAME and L-arginine on hemostatic parameters, and to reveal the effects of BPC 157 on the L-NAME- and L-arginine-induced hemostatic actions under different pathological condition: tail amputation without or with anticoagulants, heparin or warfarin.
Methods: Tail amputation, and/or i.v.-heparin (10 mg/kg), i.g.-warfarin (1.5 mg/kg/day for 3 days) were used in rats. Treatment includes BPC 157, L-NAME, L-arginine, per se and their combination.
Results: After (tail) amputation, with or without i.v.-heparin or i.g.-warfarin, BPC 157 (10 μg/kg, 10 ng/kg, i.p., i.v. (heparin), 10 μg/kg i.g. (warfarin)) always reduced bleeding time and/or haemorrhage and counteracted thrombocytopenia. As for L-NAME and/or L-arginine, we noted: L-arginine (100 mg/kg i.p.)-rats: more bleeding, less/no thrombocytopenia; L-NAME (5 mg/kg i.p.)-rats: less bleeding (amputation only), but present thrombocytopenia; L-NAME+L-arginine-rats also exhibited thrombocytopenia: L-NAME counteracted L-arginine-increased bleeding, L-arginine did not counteract L-NAME-thrombocytopenia. All animals receiving BPC 157 in addition (BPC 157 μg+L-NAME; BPC 157 μg+L-arginine, BPC 157 μg+L-NAME+L-arginine), exhibited decreased haemorrhage and markedly counteracted thrombocytopenia.
Conclusions: L-NAME (thrombocytopenia), L-arginine (increased haemorrhage) counteraction and BPC 157 (decreased haemorrhage, counteracted thrombocytopenia) with rescue against two different anticoagulants, implicate a BPC 157 modulatory and balancing role with rescued NO-hemostatic mechanisms.
PubMed Disclaimer
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
Figures Fig 1. Hematocrit-corrected platelet counts (X) before, and after bleeding period.
Hematocrit-corrected platelet counts (X) before, and after bleeding period: (a) in normal rats, BPC 157 (10 μg/kg, 10 ng/kg), L-NAME (5 mg/kg), L-arginine (100 mg/kg) given alone and/or together,applied intraperitoneally, at 30 minutes before amputation while controls received simultaneously an equivolume of saline (5ml/kg intraperitoneally); (b) in heparin (10 mg/kg intravenously) challenged rats when treated with BPC 157 (10 μg/kg, 10 ng/kg), L-NAME (5 mg/kg), L-arginine (100 mg/kg) alone or combined, intravenously, immediately after intravenous heparin while controls received simultaneously an equivolume of saline (0.5 ml/kg intravenously); (c) in warfarin rats (warfarin given intragastrically (1.5 mg/kg) once daily for 3 consecutive days, with the last challenge was at 3 hours before the bleeding procedure). BPC 157 (10 μg/kg, 10 ng/kg) was given immediately after any warfarin challenge, intragastrically while controls received simultaneously an equivolume of saline (5.0 ml/kg intragastrically). We applied L-NAME (5 mg/kg), L-arginine (100 mg/kg) alone or combined, intraperitoneally, at 30 minutes before amputation. Mean ± SD, *P<0.05 at least vs. control, 10 rats at least per group.
References
Stupnisek M, Franjic S, Drmic D, Hrelec M, Kolenc D, Radic B, et al. (2012) Pentadecapeptide BPC 157 reduces bleeding time and thrombocytopenia after amputation in rats treated with heparin, warfarin or aspirin. Thromb Res 129: 652–659. 10.1016/j.thromres.2011.07.035 - DOI - PubMed
Hrelec M, Klicek R, Brcic L, Brcic I, Cvjetko I, Seiwerth S, et al. (2009) Abdominal aorta anastomosis in rats and stable gastric pentadecapeptide BPC 157, prophylaxis and therapy. J Physiol Pharmacol 60 Suppl 7: 161–165. - PubMed
Dambisya YM, Lee TL. (1996) A thromboelastography study on the in vitro effects of L-arginine and L-NG-nitro arginine methyl ester on human whole blood coagulation and fibrinolysis. Blood Coagul Fibrinolysis 7: 678–683. - PubMed
Sakawi Y, Tarpey M, Chen YF, Calhoun DA, Connor MG, Chestnut DH, et al. (2000) Evaluation of low-dose endotoxin administration during pregnancy as a model of preeclampsia. Anesthesiology 93: 1446–1455. - PubMed
Thom SR, Fisher D, Xu YA, Garner S, Ischiropoulos H. (1999) Role of nitric oxide-derived oxidants in vascular injury from carbon monoxide in the rat. Am J Physiol 276: H984–992. - PubMed