Evaluation of antioxidative potential and cytokines mediated immunopotentiating activity of Eclipta alba (L.) Hassk. 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
Medicinal plants have gained popularity in recent era and there has been a rise in the researches related to them. They possess numerous antibacterial, antioxidants and anticancer agents which provide an alternative mode of therapy to several diseases and infections caused by drug resistant pathogenic strains, oxidative stress, etc. Eclipta alba (L.) Hassk., also known as ‘false daisy’ or ‘bhringraj’ belongs to family Aesteraceae. Since ancient times, this plant has been used in Ayurveda. The fact that human body requires additional nutrients, has given rise to consumption of eggs and meat in the country. Thus, health of poultry is important aspect as consumption of healthy poultry products will enhance the health of human population as well. On the above mentioned facts, present study was planned to explore immunomodulatory and antioxidative potential of 50% hydromethanolic extract of aerial part of Eclipta alba (EAE). Extract of the plant was prepared and evaluated for the presence of various phytoconstituents through qualitative and quantitative phytochemical analyses. DPPH and NO scavenging assays were carried out to assess the antioxidative potential of EAE. The immunomodulatory potential of EAE was evaluated through lymphocyte proliferation assay (LPA) followed by biochemical analyses (LPO, GSH, SOD, catalase and NO) in EAE treated chicken lymphocytes. Expression analysis of IL-6, IL-10 and iNOS was carried out through quantitative real time PCR in chicken lymphocytes to assess role of EAE exposure in modulating their expression. The extraction yield of EAE was found to be 8.51%. Biochemical analyses revealed the presence of various phytoconstituents. The total phenolics content and total flavonoid content in EAE was estimated to be 871.11 mg/g and 376 mg/g, respectively. Maximum non-cytotoxic doses of both, EAE and dexamethasone were determined in the lymphocytes culture system which was found to be 75μg/ml for EAE and 30ng/ml for dexamethasone. EAE displayed significant increase of 14.31% and 33.67% in T cell proliferation in case of PHA and Con A stimulated cells, respectively. There was marked increase of 30.64% in B cell proliferation in LPS stimulated cells as compared to control. The antioxidant property of the extract was revealed by DPPH assay and NO scavenging assay. The IC50 value for EAE was found to be 175.16 μg/ml in DPPH assay and 70.39 μg/ml in NO scavenging assay. There was significant decrease in the percent Lipid peroxidation and NO content after the treatment with the EAE in chicken lymphocytes. However, GSH, SOD and Catalase levels were significantly increased in the cells treated with EAE in comparison to control cells. The expression of IL-6 was found to be increased in Con A stimulated EAE treated cells, but the same was decreased in LPS stimulated EAE treated cells. iNOS expression was found to be reduced significantly in macrophages rich cells as well as in Con A and LPS stimulated EAE treated cells. However, expression of IL-10 was elevated significantly in Con A stimulated EAE treated lymphocytes but was reduced in LPS stimulated EAE treated lymphocytes. The findings of this study indicated that the hydromethanolic extract of aerial parts of Eclipta alba are rich in phytochemicals (flavonoids and phenolics). Both the extract and dexamethasone exhibited presence of antioxidantive potential and displayed immunomodulatory effects in chicken lymphocytes culture system. EAE exposure modulated the expression of IL-6, IL-10 and iNOS in chicken splenocytes. Due to the significant immunopotentiating and antioxidative potential of EAE, it could be used for development of immunoenhancing herbal preparations for poultry. However, exhaustive research is needed to characterize phytoconstituents and the findings need to be validated at protein level by employing modern biochemical and molecular biology techniques. Also the effects of Eclipta alba need to be determined on suitable in vitro and in vivo systems.
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