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Subject: Biotechnology and molecular Biology × Author: Singh, Himani × End date: 2022 × Start date: 2022 × Type: Thesis × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    In vitro evaluation of antioxidative and immunomodulatory potential of the selected medicinal plants via cytokines expression analysis in chicken splenocytes culture system
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-04) Singh, Himani; Ambwani, Sonu
    Herbal immunomodulation is being traditionally practiced in India and is gaining added importance nowadays throughout the world. Due to lack of knowledge regarding the molecular targets and underlying mechanisms involved, there is pressing need to scientifically validate the molecular mechanism behind herbal immunomodulation. Considering the above facts, four plant extracts, viz. aqueous leaf extract of Nyctanthes arbor-tristis (Night-flowering Jasmine) (NAE), hydromethanolic rhizome extract, bark extract and seeds extract of Asparagus racemosus (Shatavari) (ARE), Terminalia arjuna (Arjun) (TAE) and Trigonella foenum-graecum (Fenugreek) (TFE), respectively; were explored for their antioxidative and immunomodulatory potential in chicken splenocytes culture system. These were evaluated for the presence of various phytoconstituents through GC-MS analysis. The antioxidative potential of all the four plant extracts was evaluated by various in vitro extracts based and cell based assays. The anti-inflammatory activity of the plant extracts was determined by protein denaturation method and proteinase enzyme inhibitory assay. The MNCD of NAE, ARE, TAE, TFE and DEXA were used to give in vitro exposure in chicken splenocytes to further explore the transcriptional level modulation of the selected genes through qRT-PCR. GC-MS data analysis of NAE, ARE, TAE and TFE revealed the presence of several active phytochemicals which may be responsible for their antioxidant and immunomodulatory properties. All the four plant extracts displayed significant in vitro antioxidant and anti-inflammatory properties in various assays conducted. Expression analysis displayed significant alterations in expression of the selected genes due to in vitro exposure of NAE, ARE, TAE, TFE and DEXA. The pro-inflammatory genes were found to be downregulated while the anti-inflammatory genes were found to be significantly upregulated in the plant extracts and DEXA treated splenocytes. There was significant increase in immunostimulatory genes in the plant extracts treated splenocytes while same were found to be decreased in DEXA treated cells. There was marked reduction in the transcript level expression of APAF-1, caspase-3 and caspase-9 when treated with NAE, ARE, TAE, TFE while these genes were found to be upregulated in DEXA treated cells. From the findings of the present study, it may be concluded that NAE, ARE, TAE and TFE showed potent anti-oxidative, anti-inflammatory and immunostimulatory potential while DEXA showed anti-oxidative, anti-inflammatory and immunosuppressive property. Due to the significant immunopotentiating and antioxidative potential of the plant extracts, these could be further 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 suitable biochemical and molecular biology techniques.