IN VITRO STUDY ON PROTEASE AND THROMBOLYTIC ACTIVITY OF AQUEOUS EXTRACT FROM LEUCAS ASPERA (L.) LEAVES

  • NAGAMANI J. E. Garden city university
  • USHA SAH
  • RAVINDRANATH H. A.

Abstract

Objective: The current study is an attempt to screen for the in-vitro clot lysis and proteolytic activity of aqueous extract of Leucas aspera leaves.


Methods: Thrombolytic activity and protease activity of the crude enzyme obtained by ammonium sulfate precipitation and dialysis were assayed using blood clot and casein as substrates respectively. Native PAGE and gel documentation studies were performed to calculate the molecular weight of the enzyme.


Results: In the study, 40% salt fractioned crude enzyme sample exhibited significant thrombolytic and caseinolytic activity. Further dose dependent increased activity was observed with the maximum lytic activity of 52.11 ± 1.04 % at 1mg/ml of the sample when compared to the reference drug streptokinase (71.39 ± 0.32%).  Also 68.72 ± 0.62 U/hr of caseinolytic activity was observed for 1mg/ml of the sample fraction.


Conclusion: The study highlights and validates the efficacy of Leucas aspera leaves extract for thrombolytic and proteolytic actions. Enzyme with an approximate molecular weight, 19.89 KDa could be responsible for the significant lytic activity.


 

Keywords: Leucas aspera, Thrombolytic activity, Proteolytic activity, Native PAGE

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References

1. Kumar S, Joseph L, George M, Sharma A. A review on the anticoagulant / antithrombotic activity of natural plants used in traditional medicine. Int J Pharmaceutical Sciences Review and Research 2011;8(1):70-74.
2. Irfan Newaz Khan, Md. Razibul Habib, Md. Mominur Rahman. Thrombolytic potential of Ocimum sanctum L., Curcuma longa L., Azadirachta indica L. and Anacardium occidentale L. J Basic and Clinical Pharmacy 2011;002 (003): 125 -127.
3. Collen D. Coronary thrombolysis: Streptokinase or recombinant tissue-type plasminogen activator. Ann Intern Med 1990;112 (7): 529-38.
4. Sweta Prasad, Rajpal S Kashyap, Jayant Y Deopujari, et al. Development of an In-vitro model to study clot lysis activity of thrombolytic drug. Thrombosis Journal 2006; 4:1-14.
5. Nicolini F A, Nichols W W, Mehta J L, Saldeen T G, et al. Sustained reflow in dogs with coronary thrombosis with K2P, a novel mutant of tissue plasminogen activator. J Am Coll Cardiol 1992; 20: 228-235.
6. Adams D S, Griffin L A, Nachajko W R, Reddy V B, Wei C M. A synthetic DNA encoding a modified human urokinase resistant to inhibition by the serum plasminogen activator inhibitor. J Biol Chem 1991;266: 8476-8482.
7. Lijnen H R, Vanhoef B, DeCock F, Okada K, et al. On the mechanism of fibrin-specific plasminogen activation by staphylokinase. J Biol Chem 1991;266: 11826-11832.
8. Gesler WM. Therapeutic landscapes: medical issues in light of the new cultural geography. Soc Sci Med 1992;34 (7): 735–746.
9. Yamamoto J, Yamada K, Naemura A, Yamashita T, Arai R. Testing various herbs for antithrombotic effect. Nutrition 2005; 21 (5): 580-587.
10. Prasad S, Kashyap R S, Deopujari J Y, Purohit H J, Taori G M, Daginawala H F. Effect of Fagonia Arabica (Dhamasa) on In vitro thrombolysis. BMC Complementary and Alternative Medicine 2007;7: 36: 1-6.
11. Ai Lan Chew1, Jeyanthi James Antony Jessica, Sreenivasan Sasidharan. Asian Pacific J Tropical Biomedicine 2012;2 (3) 176 – 180.
12. Goudgaon N M, Basavaraj N R, Vijayalaxmi A. Antiinflammatory activity of different fractions of Leucas aspera. Indian J Pharmacol 2003;35:397–398.
13. Srinivas K, Rao MEB, Rao S S. Anti-inflammatory activity of Heliotropium indicum Linn and Leucas aspera Spreng in albino rats. Indian J Pharmacol 2000;32:37–38.
14. Sadhu S K, Okuyama E, Fujimoto H, Ishibashi M. Separation of Leucas aspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. Chem Pharm Bull 2003;51 (5): 595–598.
15. Sadhu S K, Okuyama E, Fujimoto H, Ishibashi M. Diterpenes from Leucas aspera inhibiting prostaglandin-induced contractions. J Nat Prod 2006;69(7):988–94.
16. Chew A L, Jessica J J, Sasidharan S. Antioxidant and antibacterial activity of different parts of Leucas aspera. Asian Pac J Trop Biomed 2012;2 (3): 176–80.
17. Murata J, Satake M, Suzuki T. Studies on snake venom XII distribution of proteinase activities among Japanese and Formosan snake venoms. J Biochem 1963; 53:431 43.
18. Neurath H. The diversity of proteolytic enzymes. A Practical Approach. Oxford, UK: IRL Press. 1990;1–12.
19. Kunwar Awaneesh Singh, Manasa K Nayak, Medicherla V Jagannadham, Debabrata Dash. Thrombolytic along with the anti-platelet activity of crinumin, a protein constituent of Crinum asiaticum. Blood Cells, Molecules, and Diseases 2011;47 (2), 129-32.
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J. E., N., U. SAH, and R. H. A. “IN VITRO STUDY ON PROTEASE AND THROMBOLYTIC ACTIVITY OF AQUEOUS EXTRACT FROM LEUCAS ASPERA (L.) LEAVES”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 13, no. 7, May 2021, doi:10.22159/ijpps.2021v13i7.41210.
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