This study aims to investigate whether tested organic extracts possess antithrombotic properties with minimal or no toxicity and to predict the activity of some of their isolated compounds.
An in vitro thrombolytic model was used to check the clot lysis effect of four Bangladeshi herbal extracts viz., roots of Curculigo recurvata W.T. Aiton (Satipata), leaf of Amorphophallus bulbifer Roxb. (Olkachu), leaf of Phyllanthus sikkimensis Muell. Arg., and whole plant of Thunbergia grandiflora Roxb. (Nillata) using streptokinase as a positive control and water as a negative control. Cytotoxicity was screened by brine shrimp lethality bioassay using vincristine sulfate as positive control. In silico prediction of activity spectra for substances (PASS) prediction was applied for phytoconstituents, namely, nyasicoside, glucomannan, grandifloric acid, serine, and alanine.
Using an in vitro thrombolytic model, C. recurvata, A. bulbifer, P. sikkimensis, and T. grandiflora showed 28.10±1.64%, 42.47±1.96%, 32.86±1.92%, and 25.51±1.67% of clot lysis, respectively. Reference drug streptokinase exhibited 75.00±3.04% clot lysis. Examined herbs showed significant (p<0.001) percentage (%) of clot lysis compared to negative control. In brine shrimp cytotoxic assay, C. recurvata, A. bulbifer, P. sikkimensis, and T. grandiflora showed LC50 values 210.64±3.44, 98.51±1.47, 187.29±2.01, and 386.43±3.02 μg/mL, respectively, with reference to vincristine sulfate (LC50 0.76±0.04). PASS predicted that examined phytoconstituents have a wide range of biological activity.
Through our study it was found that A. bulbifer and P. sikkimensis could be considered as very promising and beneficial thrombolytic agents.
The authors are grateful to the authority of International Islamic University Chittagong, Bangladesh, for providing the facilities to conduct this research work. The authors are thankful to all members of GUSTO (a research group) for their kind help in the experiment. The authors are also thankful to the Taxonomist and Professor, Dr. Shaikh Bokhtear Uddin, Department of Botany, University of Chittagong, for identifying all the plants.
Author contributions: All the authors have accepted responsibility for the entire contents of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
5. Nicolini FA, Nichols WW, Mehta JL, Saldeen TG, Schofield R, Ross M, et al. Sustained reflow in dogs with coronary thrombosis with K2P, a novel mutant of tissue-plasminogen activator. J Am Coll Cardiol 1992;20:228–35.10.1016/0735-1097(92)90164-ISearch in Google Scholar
6. Demrow HS, Slane PR, Folts JD. Administration of wine and grape juice inhibits in vivo platelet activity and thrombosis in stenosed canine coronary arteries. Circulation 1995;91:1182–8.10.1161/01.CIR.91.4.1182Search in Google Scholar
8. Leta GC, Mourão PA, Tovar AM. Human venous and arterial glycosaminoglycans have similar affinity for plasma low-density lipoproteins. Biochim Biophys Acta 2002;1586:243–53.10.1016/S0925-4439(01)00102-8Search in Google Scholar
9. Li Z, Wang H, Li J, Zhang G, Gao C. Basic and clinical study on the antithrombotic mechanism of glycosaminoglycan extracted from sea cucumber. Chin Med J 2000;113:706–11.Search in Google Scholar
10. Rajapakse N, Jung WK, Mendis E, Moon SH, Kim SK. A novel anticoagulant purified from fish protein hydrolysate inhibits factor XIIa and platelet aggregation. Life Sci 2005;76:2607–19.10.1016/j.lfs.2004.12.010Search in Google Scholar
14. Majumdar K, Datta B. A study on ethnomedicinal usage of plants among the folklore herbalists and Tripuri medical practitioners: Part-II. Nat Prod Radiance 2007;6:66–73.Search in Google Scholar
15. Chifundera K, Palazzino G, Messana I, Ping L, Galeffi C, Cannarsa G. Norlignan glucosides from Curculigo recurvata. Phytochemistry 1994;35:1343–8.10.1016/S0031-9422(06)80122-6Search in Google Scholar
16. Reddy SK, Kumar SA, Kumar VD, Ganapaty S. Anti-inflammatory and analgesic activities of Amorphophallus bulbifer (Roxb) Kunth whole plant. Trop J Pharm Res 2013;11:971–6.10.4314/tjpr.v11i6.14Search in Google Scholar
17. Shete C, Wadkar S, Inamdar F, Gaikwad N, Patil K. Antibacterial activity of Amorphophallus konkanensis and Amorphophallus bulbifer tuber. Asian J Pharm Clin Res 2015;8:98–102.Search in Google Scholar
18. Galeffi C, Nicoletti M, Palazzino G, Federici E. Hypoxidaceae, a monocotyledons family source of norlignan glucosides with different biological activity. Trad Med Mat Medica 2002;1:121–8.Search in Google Scholar
19. Morshed AJ. A survey of medicinal plants as regards to their uses by the tribal practitioners in the Chittagong hill tracts of Bangladesh to check various diseases. Hamdard Med 2013;56:18–40.Search in Google Scholar
21. Filimonov DA, Lagunin AA, Gloriozova TA, Rudik AV, Druzhilovskii DS, Pogodin PV, et al. Prediction of the biological activity spectra of organic compounds using the PASS online web resource. Chem Heterocycl Compd 2014;50:444–57.10.1007/s10593-014-1496-1Search in Google Scholar
22. Prasad S, Kashyap RS, Deopujari JY, Purohit HJ, Taori GM, Daginawala HF. Development of an in vitro model to study clot lysis activity of thrombolytic drugs. Thromb J 2006;4:1–4.10.1186/1477-9560-4-14Search in Google Scholar
23. Meyer B, Ferrigni N, Putnam J, Jacobsen L, Nichols DJ, McLaughlin J. Brine shrimp: a convenient general bioassay for active plant constituents. Planta Medica 1982;45:31–4.10.1055/s-2007-971236Search in Google Scholar
24. Deng LL, Wu Y, Qin QJ, Wang Q, Zhong G. Formulation optimization of thermosensitive edible film based on modified konjac glucomannan from Amorphophallus bulbifer. Food Sci 2013;34:113–9.Search in Google Scholar
26. Nair A, Subramanian S, Nagarajan S. Chemical composition of nectar in Thunbergia grandiflora. Curr Sci 1964;401:1.Search in Google Scholar
28. Khurana N, Ishar MP, Gajbhiye A, Goel RK. PASS assisted prediction and pharmacological evaluation of novel nicotinic analogs for nootropic activity in mice. Eur J Pharmacol 2011;662:22–30.10.1016/j.ejphar.2011.04.048Search in Google Scholar PubMed
29. Tiwari P, Kumar B, Kaur M, Kaur G, Kaur H. Phytochemical screening and extraction: a review. Int Pharm Sci 2011;1: 98–106.Search in Google Scholar
31. Liu S, Manson JE, Lee IM, Cole SR, Hennekens CH, Willett WC, et al. Fruit and vegetable intake and risk of cardiovascular disease: the Women’s Health Study. Am J Clin Nutr 2000;72:922–8.10.1093/ajcn/72.4.922Search in Google Scholar
32. Bazzano LA, He J, Ogden LG, Loria CM, Vupputuri S, Myers L, et al. Fruit and vegetable intake and risk of cardiovascular disease in US adults: the first National Health and Nutrition Examination Survey Epidemiologic Follow-up Study. Am J Clin Nutr 2002;76:93–9.10.1093/ajcn/76.1.93Search in Google Scholar
33. Torres-Urrutia C, Guzman L, Schmeda-Hirschmann G, Moore-Carrasco R, Alarcon M, Astudillo L, et al. Antiplatelet, anticoagulant, and fibrinolytic activity in vitro of extracts from selected fruits and vegetables. Blood Coagul Fibrinolysis 2011;22:197–205.10.1097/MBC.0b013e328343f7daSearch in Google Scholar
34. Yuri JA, Neira A, Quilodran A, Motomura Y, Palomo I. Antioxidant activity and total phenolics concentration in apple peel and flesh is determined by cultivar and agroclimatic growing regions in Chile. J Food Agric Environ 2009;7:513–7.Search in Google Scholar
35. Kowalski M, Brown G, Bieniasz M, Oszajca K, Chabielska E, Pietras T, et al. Cloning and expression of a new recombinant thrombolytic and anthithrombotic agent – a staphylokinase variant. Acta Biochim Pol 2009;56:41.10.18388/abp.2009_2515Search in Google Scholar
36. Pantzar M, Ljungh Å, Wadström T. Plasminogen binding and activation at the surface of Helicobacter pylori CCUG 17874. Infect Immun 1998;66:4976–80.10.1128/IAI.66.10.4976-4980.1998Search in Google Scholar
37. Parry MA, Zhang XC, Bode W. Molecular mechanisms of plasminogen activation: bacterial cofactors provide clues. Trends Biochem Sci 2000;25:53–9.10.1016/S0968-0004(99)01521-2Search in Google Scholar
39. Rahman MA, Imran TB, Islam S. Antioxidative, antimicrobial and cytotoxic effects of the phenolics of Leea indica leaf extract. Saudi J Biol Sci 2013;20:213–25.10.1016/j.sjbs.2012.11.007Search in Google Scholar PubMed PubMed Central
41. Fowles RG, Mootoo BS, Ramsewak RS, Khan A. Toxicity-structure activity evaluation of limonoids from Swietenia species on Artemia salina. Pharm Biol 2012;50:264–7.10.3109/13880209.2011.581670Search in Google Scholar PubMed
42. Okwuosa V, Molta B, Ebele S. Toxicity of aqueous bark extract of the tree Balanites aegyptiaca on the fish Oreochromis niloticus. Appl Parasitol 1993;34:89–94.Search in Google Scholar
43. Parra AL, Yhebra RS, Sardiñas IG, Buela LI. Comparative study of the assay of Artemia salina L. and the estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant extracts. Phytomedicine 2001;8: 395–400.10.1078/0944-7113-00044Search in Google Scholar PubMed
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