Journal of Environment and Sociobiology

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Effect of Partially Purified Fraction from the Crude Aqueous Extract of Allium sativum L. on Sperm Immobilization


Affiliations
1 Department of Zoology, Charuchandra College, 22-Lake Road, Kolkata-700 029, West Bengal, India
     

   Subscribe/Renew Journal


To elucidate the nature and the probable mode of action of the partially purified in vitro human sperm immobilizing fraction of Allium sativum L., with a view to develop a herbal intravaginal spermicidal agent partial purification was carried out using different types of chromatography. After each step of purification, the purified fractions were tested on human ejaculated sperm to verify the instant immobilization activity and thus the most active fraction underwent further purification. For characterization of its nature, Nuclear magnetic spin resonance (NMR), Gas Liquid Chromatography (GLC) and Electron spray ionization mass spectroscopy (ESI-MS) were carried out. In order to understand the mechanism of action, several sperm functional parameters such as viability, Hypo-Osmotic Swelling (HOS), 5´ nucleotidase, acrosin and acrosome intactness were estimated when the sperm cells were incubated with the partially purified extract. Characterization showed that the partially purified fraction contained β-linked heptasaccharide consisting of glucose molecule as aglycone part, whose approximate molecular weight was 1152. The minimum inhibitory concentration (MIC) and Effective Concentration 50 (EC50) of this fraction was 50 mg/ml and 16 mg/ml, respectively. When treated sperm was compared with the control, there was a loss of more than 65% in the activity of plasma membrane marker enzyme 5’ nucleotidase, 55% loss in sperm viability, 78% reduction in HOS 80% decrease in the activity of acrosin and acrosome intactness. Results revealed the in vitro sperm immobilization activity of the partially purified extract, consisting of glucose molecules as one of the major constituents, which exert its effect mainly on plasma and acrosomal membrane architecture.

Keywords

Aqueous Extract, Partial Purification, Characterization, Human Sperm, Sperm Immobilization, Intra Vaginal Contraceptive.
Subscription Login to verify subscription
User
Notifications
Font Size


  • Bhattacharyya, A. K. and Kanjilal, S. 2003. Assessment of sperm functional competence and sperm egg interaction. Mol. Cell Biochem., 253(1-2): 255-261.

  • Chakrabarti, K., Pal, S. and Bhattacharyya. A. K. 2003. Sperm immobilization activity of Allium sativum (L.) and other plant extracts. Asian J. Androl., 5(2): 131-135.

  • Chaudhury, K., Bhattacharyya, A. K. and Guha, S. K. 2004. Studies on the membrane integrity of human sperm treated with a new injectable male contraceptive. Hum. Reprod., 19(8): 1826-1830.

  • Dixit, V.P. and Joshi, S. 1982. Effect of chronic administration of garlic (Allium Linn.) on testicular function. Indian J. Exp. Biol., 20(7): 534-536.

  • O’Gara, E. A., Hill, D. J., Maslin, D. J. 2000. Activities of garlic oil, garlic powder, and their diallyl constituents against Helicobacter pylori. Appl. Environ. Microbiol., 66(5): 2269-2273.

  • Gopakrishnan, K. 1995. Standardized procedures in human semen analysis. Curr. Sci., 68(4): 353-362.

  • Lohiya, N. K., Pathak, N., Mishra, P. K. and Manivannam, B. 2000. Contraceptive evaluation and toxicological study of aqueous extract of the seeds of Carica papaya in male rabbits. J. Ethnopharmacol., 70(1): 17-27.

  • Pakrashi, A., Ray, H., Pal, B. C. and Mohato, S. B., 1991. Sperm immobilizing effect of triterpene saponins from Acacia auriculiformis. Contraception, 43(5): 475-483.

  • Qian, Y. X., Shen, P. J., Xu, R. Y., Liu, G. M., Yang, et al. 1986. Spermicidal effect in vitro by the active principle of garlic. Contraception, 34(3): 295-302.

  • Rajasekaran, M., Nair, A. G. R., Hellstrom, W. J. G., Sikka, S. C. 1993. Spermicidal activity of an antifungal saponin obtained from the tropical herb Mollugo pentaphylla. Contraception, 47(4): 401-412.

  • Talwar, G. P., Garg, S., Dhar, V., Chabra, R., Ganju, A. et al. 1995. Praneem polyherbal cream and pessaries with dual properties of contraception and alleviation of genital infections. Curr. Sci., 68(4): 437-440.

  • WHO. 1999. Laboratory Manual for the Examination of Human Semen and Sperm-cervical Mucus Interaction. 4th ed. Cambridge: Cambridge University Press, pp. 1-125.

  • Zaneveld, L. J. D. 1982. The epididymes. In: L. J. D. Zaneveld and R. T. Chatterton (eds.), Biochemistry of Mammalian Reproduction. John Wiley and Sons, NY.


Abstract Views: 132

PDF Views: 0




  • Effect of Partially Purified Fraction from the Crude Aqueous Extract of Allium sativum L. on Sperm Immobilization

Abstract Views: 132  |  PDF Views: 0

Authors

Kausiki Chakrabarti
Department of Zoology, Charuchandra College, 22-Lake Road, Kolkata-700 029, West Bengal, India

Abstract


To elucidate the nature and the probable mode of action of the partially purified in vitro human sperm immobilizing fraction of Allium sativum L., with a view to develop a herbal intravaginal spermicidal agent partial purification was carried out using different types of chromatography. After each step of purification, the purified fractions were tested on human ejaculated sperm to verify the instant immobilization activity and thus the most active fraction underwent further purification. For characterization of its nature, Nuclear magnetic spin resonance (NMR), Gas Liquid Chromatography (GLC) and Electron spray ionization mass spectroscopy (ESI-MS) were carried out. In order to understand the mechanism of action, several sperm functional parameters such as viability, Hypo-Osmotic Swelling (HOS), 5´ nucleotidase, acrosin and acrosome intactness were estimated when the sperm cells were incubated with the partially purified extract. Characterization showed that the partially purified fraction contained β-linked heptasaccharide consisting of glucose molecule as aglycone part, whose approximate molecular weight was 1152. The minimum inhibitory concentration (MIC) and Effective Concentration 50 (EC50) of this fraction was 50 mg/ml and 16 mg/ml, respectively. When treated sperm was compared with the control, there was a loss of more than 65% in the activity of plasma membrane marker enzyme 5’ nucleotidase, 55% loss in sperm viability, 78% reduction in HOS 80% decrease in the activity of acrosin and acrosome intactness. Results revealed the in vitro sperm immobilization activity of the partially purified extract, consisting of glucose molecules as one of the major constituents, which exert its effect mainly on plasma and acrosomal membrane architecture.

Keywords


Aqueous Extract, Partial Purification, Characterization, Human Sperm, Sperm Immobilization, Intra Vaginal Contraceptive.

References