Determination of antioxidative potential and cytokines mediated immunomodulation due to in vitro exposure of Aegle marmelos (l.) corr. in chicken lymphocytes culture system

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Date
2018-08
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Traditional medicine system has always been a part of different cultures and civilizations around the world. India has one of the richest plant based medicine system. Poultry system is vulnerable to new evolving bacterial, viral strains and different environmental stresses which result in huge economic loss. Aegle marmelos (L.) Corr. belongs to Rutaceae family, commonly known as “bael” in Hindi. Various extracts of Aegle marmelos are reported to have antiproliferative activity in splenocytes and peripheral blood mononuclear cells. On the basis of above mention lines, the present study was planned to explore the immunomodulatory and antioxidative potential of aqueous extract of leaves of Aegle marmelos (AME) in chicken lymphocytes. To check the presence of various phytoconstituents in AME, various qualitative and quantitative phytochemical analyses were carried out. Further, two cytokines (IL-6, IL-10) and iNOS expression analysis in AME exposed chicken lymphocytes was carried out through quantitative real time PCR. The immunomodulatory potential was explored through lymphocyte proliferation assay, while DPPH assay, NO radical scavenging assay and various cell based assays viz. LPO, SOD, Catalase, Reduced GSH and Nitric oxide estimation were conducted to explore antioxidative potential of AME. The extraction yield of aqueous extract of Aegle marmelos (AME) was found to be 14.1%. In biochemical analysis, the extract showed presence of various phytoconstituents. Total phenolic and flavanoids contents in AME was estimated to be 480 GAE mg/g and 371.2 RE mg/g respectively. The maximum non cytotoxic dose (MNCD) of extract was determined to be 75μg/ml in chicken lymphocyte culture system. AME showed immunosuppressive potential by decreasing the proliferation of T and B lymphocytes in mitogen stimulated cells. The percent decrease of 6.20% and 15.5% % in T cell proliferation was observed in case of Con A and PHA stimulated cells respectively. Significant decrease of 9.23% in B cell proliferation was observed due to AME exposure in case of LPS stimulated cells as compared to control. AME showed significant antioxidative activity in DPPH and Nitric oxide radical scavenging assays with IC50 value for DPPH and NO scavenging assays of 233.3 μg/ml and 113.80μg/ml, respectively. After the exposure of the extract in lymphocytes, the level of membrane lipid peroxidation and nitric oxide content was decreased. The level of reduced GSH, SOD and catalase was increased in chicken lymphocytes after treatment with AME. In quantitative real time PCR, the expression level of proinflammatory cytokine IL-6 and inducible nitric oxide synthase was significantly decreased in comparison to control but the expression of anti-inflammatory cytokine IL-10 was significantly increased after the exposure of AME at MNCD. Thus, it could be concluded that AME displayed significant antioxidative and immunosuppressive potential in chicken lymphocytes culture system which could be correlated with the alteration in IL-6, IL-10 and iNOS expression levels. However, there is need for further suitable in vitro and in vivo analyses to exploit this plant resources based therapeutic preparation for certain disease conditions.
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