P105 Effects of Azardirchata indica and Acatia nilotica extracts on splenocytes proliferation and IFN-Γ cytokine induction in wistar albino rats

Abstract

Introduction One of the therapeutic strategies in Ayurvedic medicine is to increase body’s natural resistance to the disease causing agent rather than directly neutralizing the agent it self. This concept in modern scientific understanding would mean enhancement of immune responsiveness of an animal against pathogens by nonspecifically activating the immune system using immunomodulatory agents of plant origin. Use of herbal medicines, as immunomodulators, which alter the activity of immune function through the dynamic regulation of informational molecules such as cytokines. Modulation of cytokines’ secretions may offer novel approaches in the treatment of variety of diseases.This study was designed to investigate the effect of A.indica and A.nilotica on cytokine induction and splenocyte proliferation to establish the concept. Methods MDBK Cell line was already maintained in the Animal Cell Culture facility under Niche Area of Excellence Project, Department of Epidemiology, DUVASU, Mathura. The cell lines were grown in RPMI 1640 and incubated at 37 °C in 5% CO2.The animals were obtained from Animal House facility at DUVASU, Marthura. The spleens were aseptically removed, and placed in RPMI 1640. Single cell suspensions were prepared and splenocytes were cultured with sample and mitogen or without mitogen for specific cell differentiation to T and B lymphocytes (Asia et al.). The cell proliferation was measured by MTT assay.Cell proliferation was expressed as percentage of viable cells of treated samples to control samples. Concentration of IFN-Gamma was measured using the commercially available ELISA kits according to the protocol provided by the manufacturer. Data processsed for statistical analysis using student t-test. Results In vitro Effect of aq. Extract of A.indica and A.nilotica leaves on splenocyte proliferation: The mean value for splenocyte proliferation using four different concentration (62.5, 125, 250 & 500 μg/well) of A. indica were 0.120 ± 0.001, 0.115 ± 0.004, 0.089 ± 0.005 & 0.073 respectively The mean value of control was 0.112 ± 0.002. The result showed the maxium inhibition in splenocyte proliferation in spleen cells harvested from normal mice treated with 250 μg/well and 500 μg/well A. indica extract which were 20.53% and 34.82% respectively. While as The mean value for splenocyte proliferation using four different concentration (62.5, 125, 250 & 500 μg/well) of A. nilotica were 0.085 ± 0.002, 0.054 ± 0.001, 0.015 ± 0.001 and 0.013 ± 0.00 respectively. The mean value of control was 0.112 ± 0.002. In-vitro Effect of aq. Extract of A.indica and A.nilotica leaves on induction of IFN-Γ cytokine. The mean value for IFN-Γ secretion in presence of four different concentration (62.5, 125, 250 & 500 μg/well) of A. indica were 5.912 ± 4.181,5.117 ± 3.618, 4.653 ± 3.290 & 2.681 ± 1.896 respectively while in ConA control group mean value was 2.692 ± 1.905. The maximum reduction was noticed with 250 & 500 μg/well of A. indica extractwhich were 22.21% and 29.56% respectively. while as The mean value for IFN-Γ secretion in presence of four different concentration (62.5, 125, 250 & 500 μg/well) of A. nilotica were 4.630 ± 3.274, 4.609 ± 3.259, 1.596 ± 1.128 & 0.127 ± 0.089 respectively while in ConA control group mean value was 2.692 ± 1.905. Conclusion Work suggests the involvement of A. nilotica and A. indica in modulating the cytokines and affecting splenocyte proliferation.