M Rinku, V Prasanth, G Parthasarathy
immunostimulant, phagocytosis, urena lobata linn
M Rinku, V Prasanth, G Parthasarathy. Immunomodulatory activity of the methanolic extract of Urena lobata Linn.. The Internet Journal of Pharmacology. 2008 Volume 7 Number 1.
Immunology is thus probably one of the most rapidly developing areas of biomedical research and has great promises with regard to the prevention and treatment of a wide range of disorders. Inflammatory diseases of the skin, gut, respiratory tract, joints and central organs In addition, infectious diseases are now primarily considered immunological disorders, while neoplastic diseases and organ transplantation and several autoimmune diseases may involve in an immunosuppressive state The immune system is known to be involved in the etiology as well as the path physiologic mechanism of many diseases
Macrophages are a corn stone of the innate immune system. White blood cells are the mainstay of immune system. Some white blood cells known as macrophage play a function in innate immunity by surrounding ingesting and destroy invading bacteria and other foreign organisms in a process called phagocytosis. Macrophages play an important role in adaptive immunity in that they attach to invading antigens and deliver them to destroy by other components of adaptive immunity.Phagocytosis is the cellular process of engulfing solid particles by the cell membrane to form phagosome or “food vacuole”. Phagocytosis is involved in the acquisition of nutrients for some cells, and in the immune system is a major mechanism used to remove pathogens and cell debris; bacteria, dead tissue cell and small mineral particles are all examples of objects that may be phagocytosed.
Neutrophil granulocytes usually make up to 80% of white blood count. They have multilobed nuclei and lightly staining granules. They assist in destruction of foreign particles by immune system. The use of variety of agents to enhance immunological and non-specific host defenses and thus to modify the defenses and thus to modify the defense favorable is an exciting development in immunopharmacology.Such agents may act by increasing the humoral antibody responses, by enhancing the phagocytic activity of macrophages or by modifying the cell mediated immune response 5 Thus in our present study, we have attempted to evaluate the immunomodulatory potency to the methanolic extract of
Materials And Methods
The freshly collected plant was washed and sun-dried and coarsely powdered until able to pass through sieve number 40
Preparation of Extract
The standardized coarse powder of plant material was subjected to soxhlet extraction using methanol as the solvent. The dark green filtrate obtained was concentrated. The crude methanolic extract was used for the presence studies and the extract also subjected to preliminary phytochemical investigations.
Preparation of the test sample
Sample for invitro study was prepared by dissolving 2.0gm of crude extract in 20ml of distilled water and to obtain a solution of 100mg/ml. From this stock solution, different working dilution were prepared to get concentration of 5,10,20,50,100µg/ml. Neutrophils of the blood withdrawn from healthy human volunteers were used to study the activity. Normal saline was used as control
Preparation of the Phosphate Buffer Solution (PBS)
2.38 gm of disodium hydrogen phosphate was dissolved in 0.19gm of potassium dihydrogen phosphate and 8gm of sodium chloride and made up to 1000ml with distilled water and pH was adjusted to 7.4.
Preparation of suspension
Candida albicans suspension was incubated in the Sabradous broth for 3-7 days. After incubation the cells were centrifuged and the supernatant was discarded. Pellet was re –suspended in PBS and centrifuged again for 3-4 times. The final cell button was re-suspended in PBS and adjusted to a concentration of 2 x10 6 /ml
Human blood (2 drops) obtained by finger prick method was collected on to a sterile glass slide. The slide was kept on a cotton pad in a sterile Petri plate and incubated at 37°Cfor 25 minutes. After incubation the clot was removed very gently and the slide was slowly drained with sterile normal saline, taking care not wash the adhered neutrophils (invisible).The slide was flooded with pre determined concentrations (5-100µg/ml) of sample and incubated at 37°C for onre hour. The slide was drained, fixed with suspension of Candida albicans and incubated at 37°C for one hour. The slide drained, fixed with methanol and stained with Giesma stain. Positive control was tested by preparing the slide in the same way with pooled with normal human blood.
Evaluation of Phagocytosis
The mean number of
Results And Discussion
Immunomodulatory agents of plant and animal origin increase the immune responsiveness of the body against pathogens by activating the non-specific immune system. However; there is a need to subject such medicinal plants to systematic studies to substantiate the therapeutic claims made with regard to their clinical utility 8 .
Recently, there is an enthusiasm towards exploration of a novel group of compounds from natural sources that modulate the immune response of living systems and influence the disease process. 9
In the present study, the preliminary phytochemical investigation on
The effect of methanolic extracts of
Effect of the methanolic extract of
From the results obtained, it can be concluded that, methanolic extract of
The authors wish to thanks Mrs.Kavitha (chairperson) and Mrs. Anitha Prasad (member of management) of Gautham College of Pharmacy, Bangalore, India for providing facilities to carry out the research work