• B. Manjula University of Mysore
  • Y. H. Mahadeswara Swamy University of Mysore
  • S, Devaraja Tumkur University
  • K. S. Girish Tumkur University
  • K. Kemparaju Department of Studies in Biochemistry, University of Mysore, Manasagangothri, Karnataka, Mysore 570006, India


Objective: To investigate the biochemical events that are associated with the skin softening, cleansing and wound healing properties of the cucumber (Cucumis sativus L) sap extract.

Methods: Preparation of cucumber sap extract (CSE). Assay of CSE for proteolytic activity, plasma re-calcification time, APTT, PT, thrombin-like activity, plasmin-like activity, and effect on platelet aggregation and wound healing property by physical, biochemical and histological examinations. Appropriate positive and negative controls were maintained wherever necessary.

Results: CSE decreased the plasma re-calcification time and prothrombin time (PT) and showed factor VII (pro-convertin) like activity. EGTA or EDTA pre-treated CSE did not alter the plasma recalcification time and PT. CSE readily hydrolyzed the plasma clot and azocasein; while, IAA pre-treated CSE did not hydrolyze the plasma clot and azocasein. CSE inhibited the agonists collagen, ADP and epinephrine induced platelet aggregation in PRP in the order epinephrine>collagen>ADP with the respective IC50 of 22 ± 2.5, 20 ± 3 and 11 ± 2 µg/ml. PMSF pre-treated but not IAA and EDTA pre-treated CSE lost the platelet aggregation inhibition property. Further, CSE augmented wound healing process including the scar removal in a mouse model. The SOD, CAT, GSH activities and hydroxyproline, hexosamine and hexuronic acid contents were increased while, NO, LPO and MPO activities were decreased compared to control values. Histological study revealed accelerated wound healing involving epithelialisation and re-formation of skin following CSE treatment compared to Neosporin.

Conclusion: CSE contain metallo-, serine and cysteine proteases, and interfere in clot formation, dissolution and wound healing process, which validates the use of cucumber as cosmetics and to treat wounds by traditional healers.


Keywords: Cucumber sap extract (CSE), Hemostasis, fibrin clot dissolution, Ethylene diamine tetra acetic acid (EDTA), Phenyl methyl sulphonyl fluoride (PMSF), Iodoacetic acid (IAA), Metalloprotease, Serine protease, Cysteine protease, Wound healing


Download data is not yet available.


1. Stadelmann WK, Digenis AG, Tobin GR. Physiology and healing dynamics of chronic cutaneous wounds. Am J Surg 1998;176:26-38.
2. Rajinder R, Shahid P, Verma PK, Pankaj NK. Medicinal plants and their role in wound healing. J Vet Online 2008;3:1-7.
3. Nema NK, Maity N, Sarkar B, Mukherjee PK. Cucumis sativus fruit-potential antioxidant, anti-hyaluronidase and anti-elastase. Arch Dermatol Res 2010;303:247-52.
4. Yamini Dixit, Anand Kar. Protective role of three vegetable peels in alloxan induced diabetes mellitus in male mice. Plant Foods Hum Nutr 2010;65:284-9.
5. Lee DH, Iwanski GB, Thoennissen NH. Cucurbitacin: ancient compound shedding new light on cancer treatment. Sci World J 2010;10:413-8.
6. Manjula B, Devaraja S, Mahadeswaraswamy YH, Girish KS, Kemparaju K. Biochemical characterization of protease isoforms in cucumber sap extract. Int J Pharm Phytopharmacol Res 2014;4:77-83.
7. Quick AJ. The prothrombin in hemophilia and obstructive jaundice. J Biol Chem 1935;107:73-85.
8. Denson KWE. Coagulation and anticoagulant actions of snake venoms. Toxicon 1969;7:5-11.
9. Chakrabarty D, Kausiki D, Antony G, Debasish B. Hemorrhagic protein of Russell’s viper venom with fibrinolytic and esterolytic activities. Toxicon 1999;38:1475-90.
10. Born GV. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962;9:927-9.
11. Frank S, Kamfer H, Dipietro LA, Burns AL. Excisional wound healing: An experimental approach. Wound Healing/Method and Protocols. Totowa, New Jersey, Humana Press; 2003. p. 3-16.
12. Angely CJ, Coppola DM. How does long-term odor deprivation affect the olfactory capacity of adult mice. Behav Brain Funct 2010;6:26.
13. Kakkar P, Das B, Viswanathan PN. A modified spectrophotometric assay of superoxide dismutase. Indian J Biochem Biophys 1984;21:130-2.
14. Aebi HU. “Catalase in methods in enzymatic analysis.” In: Methods in Enzymatic Analysis. UΗ Bergmeyer, Academic Press: New York, NY, USA; 1983;3:276–86.
15. Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem 1968;25:192-205.
16. Miranda KM, Espey MG, Wink DA. A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 2001;5:62-71.
17. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
18. Bradley PP, Christensen RD, Rothstein G. Cellular and extracellular myeloperoxidase in pyogenic inflammation. Blood 1982;60:618-22.
19. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
20. Neuman RE, Logan MA. “The determination of hydroxyproline”. J Biol Chem 1950;184:299-306.
21. Dische Z, Borenfreund E. “A spectrophotometric method for the microdetermination of hexosamines”. J Biol Chem 1950;184:517–22.
22. Bitter T, Muir HM. “A modified uronic acid carbazole reaction”. Anal Biochem 1962;4:330-4.
23. Harrison GA. Chemical methods in clinical medicine. London: Churchill. 2nd edition; 1937.
24. Jagadeesha DK, Shashidhara murthy R, Girish KS, Kemparaju K. A non-toxic anticoagulant metalloprotease: purification and characterization from Indian cobra (Naja naja) venom. Toxicon 2002;40:667-75.
25. Bamidele O, Akinnuga AM, Anyakudo MMC, Ojo OA, Ojo GB, Olorunfemi JO, et al. Haemostatic effect of methanolic leaf extract of Ageratum conyzoides in albino rats. J Med Plants Res 2010;4:2075-9.
26. Richter G, Schwarz HP, Dorner F, Turecek PL. Activation and inactivation of human factor X by proteases derived from Ficus carica. Br J Haematol 2002;119:1042–51.
27. Zhao J, Pan R, He J, Liu Y, Li DF, He RQ. Eisenia fetida protease-III-1 functions in both fibrinolysis and fibrogenesis. J Biomed Biotechnol 2007;10:1155-65.
28. Torres-Urrutia C, Guzmán L, Schmeda-Hirschmann G, Moore-Carrasco R, Alarcón M, Astudillo L, et al. Antiplatelet, anticoagulant, and fibrinolytic activity in vitro of extracts from selected fruits and vegetables. Blood Coagulation Fibrinolysis 2011;22:197-205.
29. Markland FS. Snake venoms and the hemostatic system. Toxicon 1998;36:1749-800.
30. Meier J, Stocker K. Effects of snake venoms on hemostasis. Crit Rev Toxicol 1991;21:171-82.
31. Kini RM. Serine proteases affecting blood coagulation and fibrinolysis from snake venoms. Pathophysiol Haemostasis Thromb 2005;34:200-4.
32. Patel GK, Kawale AA, Sharma AK. Purification and physicochemical characterization of a serine protease with fibrinolytic activity from latex of a medicinal herb Euphorbia hirta. Plant Physiol Biochem 2012;52:104-11.
33. Eduardo Fuentes, Luis Guzmán, Marcelo Alarcón, Rodrigo Moore, Ivan Palomo. Thrombolytic/Fibrinolytic mechanism of natural products. "Fibrinolysis and Thrombolysis", book edited by Krasimir Kolev; 2014. doi: 10.5772/57608. [Article in Press]
34. Metzig C, Grabowska E, Eckert K, Rehse K, Maurer HR. Bromelain proteases reduce human platelet aggregation in vitro, adhesion to bovine endothelial cells and thrombus formation in rat vessels in vivo. In Vivo 1999;13:7-12.
35. Singh KA, Jagannadham MV, Rao GRK, Celie PHN. Crystallization and preliminary X-ray analysis of crinumin, a chymotrypsin-like glycosylated serine protease with thrombolytic and antiplatelet activity. Acta Crystallogr 2011;67:1545-7.
36. Fronza M, Heinzmann B, Hamburger M, Laufer S, Merfort I. Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts. J Ethnopharmacol 2009;126:463-7.
37. Roy P, Amdekar S, Kumar A, Singh R, Sharma P, Singh V. In vivo antioxidative property, antimicrobial and wound healing activity of flower extracts of Pyrostegia venusta (Ker Gawl) Miers. J Ethnopharmacol 2012;140:186-92.
38. Agarwal PK, Singh A, Gaurav K, Goel S, Khanna HD, Goel RK. Evaluation of wound healing activity of extracts of plantain banana (Musa sapientum var. paradisiaca) in rats. Indian J Exp Biol 2009;47:32-40.
675 Views | 1783 Downloads
How to Cite
Manjula, B., Y. H. M. Swamy, S. Devaraja, K. S. Girish, and K. Kemparaju. “CLOT PROMOTING AND DISSOLVING PROPERTIES OF CUCUMBER (CUCUMIS SATIVUS) SAP, VALIDATING ITS USE IN TRADITIONAL MEDICINE”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 7, no. 13, Aug. 2015, pp. 104-11,