Momordica charantia seed extract exhibits strong anticoagulant effect by specifically interfering in intrinsic pathway of blood coagulation and dissolves fibrin clot

The current study explores the anticoagulant and fibrin clot-hydrolyzing properties of Momordica charantia seed extract (MCSE). MCSE hydrolyzed casein with the specific activity of 0.780 units/mg per min. Interestingly, it enhanced the clot formation process of citrated human plasma from control 146 to 432 s. In addition, the intravenous injection of MCSE significantly prolonged the bleeding time in a dose-dependent manner from control 150 to more than 800 s, and strengthened its anticoagulant activity. Interestingly, MCSE specifically prolonged the clotting time of only activated partial thromboplastin time, but not prothrombin time, and revealed the participation of MCSE in the intrinsic pathway of the blood coagulation cascade. Furthermore, MCSE completely hydrolyzed both A&agr; and B&bgr; chains of the human fibrinogen and partially hydrolyzed the &ggr; chain. However, it hydrolyzed all the chains (&agr; polymer, &agr; chain, &bgr; chain and &ggr;-&ggr; dimmers) of partially cross-linked human fibrin clot. The proteolytic activity followed by the anticoagulant effect of the MCSE was completely abolished by the 1,10-phenanthroline and phenyl methyl sulphonyl fluoride, but iodoacetic acid, EDTA, and ethylene glycol-N,N,N′,N′-tetra acetic acid did not. Curiously, MCSE did not hydrolyze any other plasma proteins except the plasma fibrinogen. Moreover, MCSE was devoid of RBC lysis, edema and hemorrhagic properties, suggesting its nontoxic nature. Taken together, MCSE may be a valuable candidate in the treatment of blood clot/thrombotic disorders.

[1]  A. Ibelli,et al.  Amblyomma americanum tick saliva serine protease inhibitor 6 is a cross‐class inhibitor of serine proteases and papain‐like cysteine proteases that delays plasma clotting and inhibits platelet aggregation , 2013, Insect molecular biology.

[2]  B. S. Vishwanath,et al.  Anti-coagulant activity of a metalloprotease: further characterization from the Indian cobra (Naja naja) venom , 2010, Journal of Thrombosis and Thrombolysis.

[3]  V. R. Devaraj,et al.  Factor Xa-like and fibrin(ogen)olytic activities of a serine protease from Hippasa agelenoides spider venom gland extract , 2009, Journal of Thrombosis and Thrombolysis.

[4]  D. Lane,et al.  The haemostatic role of tissue factor pathway inhibitor. , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[5]  B. Dhananjaya,et al.  Procoagulant activity of Calotropis gigantea latex associated with fibrin(ogen)olytic activity. , 2005, Toxicon : official journal of the International Society on Toxinology.

[6]  E. Basch,et al.  Bitter melon (Momordica charantia): a review of efficacy and safety. , 2003, American journal of health-system pharmacy : AJHP : official journal of the American Society of Health-System Pharmacists.

[7]  H. Matsuura,et al.  α-Glucosidase Inhibitor from the Seeds of Balsam Pear (Momordica charantia) and the Fruit Bodies of Grifola frondosa , 2002, Bioscience, biotechnology, and biochemistry.

[8]  K. Titani,et al.  Snake venom proteases affecting hemostasis and thrombosis. , 2000, Biochimica et biophysica acta.

[9]  J. Tainer,et al.  The food of sweet and bitter fancy , 2000, Nature Structural Biology.

[10]  Z. Wu,et al.  [Study on chemical components of Momordica charantia]. , 1998, Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials.

[11]  R. Hynes,et al.  A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  C. Sampaio,et al.  Fibrino(geno)lytic Properties of Purified Hementerin, a Metalloproteinase from the Leech Haementeria depressa , 1998, Thrombosis and Haemostasis.

[13]  C. Lans,et al.  Observations on ethnoveterinary medicines in Trinidad and Tobago. , 1998, Preventive veterinary medicine.

[14]  M. Grdiša,et al.  Fibrinolytic and anticoagulative activities from the earthworm Eisenia foetida. , 1998, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[15]  C. Lau,et al.  Anti-diabetic properties and phytochemistry of Momordica charantia L. (Cucurbitaceae). , 1996, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[16]  T. Miyata,et al.  Inhibition of serine proteases of the blood coagulation system by squash family protease inhibitors. , 1994, Journal of biochemistry.

[17]  S. Wood,et al.  Crystal structure of momordin, a type I Ribosome Inactivating Protein from the seeds of Momordica charantia , 1994, FEBS letters.

[18]  A. Cáceres,et al.  Ethnobotanical survey of the medicinal flora used by the Caribs of Guatemala. , 1991, Journal of ethnopharmacology.

[19]  H. Yeung,et al.  Effects of ginsenosides, lectins and Momordica charantia insulin-like peptide on corticosterone production by isolated rat adrenal cells. , 1987, Journal of ethnopharmacology.

[20]  R. Kini,et al.  Characterization of three edema-inducing phospholipase A2 enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid. , 1987, Toxicon : official journal of the International Society on Toxinology.

[21]  J. Collawn,et al.  Behavior of glycopolypeptides with empirical molecular weight estimation methods. 1. In sodium dodecyl sulfate. , 1980, Biochemistry.

[22]  A. Quick,et al.  A Study of the Coagulation Defect in Hemophilia and in Jaundice , 1935, Thrombosis and Haemostasis.

[23]  C. Teng,et al.  Fibrinogenolytic enzymes of Trimeresurus mucrosquamatus venom. , 1976, Biochimica et biophysica acta.

[24]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[25]  T. Suzuki,et al.  Studies on snake venom. XIII. Chromatographic separation and properties of three proteinases from Agkistrodon halys blomhoffii venom. , 1963, Journal of biochemistry.

[26]  H. Ikezawa,et al.  Studies on the quantitative method for determination of hemorrhagic activity of Habu snake venom. , 1960, Japanese journal of medical science & biology.

[27]  H. G. Boman,et al.  Chromatography of rattlesnake venom; a separation of three phosphodiesterases. , 1957, Biochimica et biophysica acta.

[28]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.