Protective effect of Vitex nigundo on Freunds complete adjuvant induced arthritis by regulating antioxidant and inflammatory mediators

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3.1. Phytochemical analysis Phytochemical analysis of VNE revealed the presence of TPC, TFC and TTC. Total phenolic content of VNE was 4.52 ± 0.04 mg GAE/g, total triterpenoid content 18.10 ± 0.11 mg UAE /g and total flavanoid content 5.38 ± 0.04 mg QHE/g of test sample (Table 1) 3.2. Free radical scavenging activity (FRSA) VNE showed FRSA by donating the hydrogen atom or electron to stable free radical DPPH. A linear relationship was observed in the DPPH radical scavenging activity and VNE concentrations (Fig 1a). The IC50 value for BHT and VNE were 29.51 ± 0.74 µg and 2.07 ± 0.05 mg respectively. 3.3. Metal chelating activity Metal binding capacity was found to increase with increasing concentrations of VNE in a linear manner (Fig 1b). EDTA showed IC 50 value of 26.68 ± 0.39 whereas it was 2.51 ± 0.03 mg for VNE. 3.4. Reducing Power A concentration dependent increase in reducing ability was observed with VNE (Fig 1c). 3.5. In vitro DNA damage The result of the plasmid nicking assay showed hydroxyl radical induced DNA damage by transforming its native supercoiled form (Lane 1) to open circular or linear form (Lane 2). (Fig 2). In the presence of BHT (50 µg, Lane 3) and VNE (Lane 4 and Lane 5; 25 and 50 µg) the native supercoiled form was retained. VNE showed a concentration dependent increase in supercoiled form. 3.6. Acute Toxicity Test and dose selection The result of acute toxicity tests showed no remarkable changes in behavior of any group of animals at all the selected doses (1.0, 2.0, 3.0 and 5.0 g/kg body weight, p.o.) as compared with the control. No mortality was observed till 72 hrs of dose administration. No significant changes were observed in liver and kidney function test in serum (Table 2). Histo... ... middle of paper ... ... to control (Group 1, Table 5) however the decrease was insignificant (p > 0.05) in erythrocytes. Indomethacin (p < 0.001) and VNE (p < 0.01) both improved GSH level in the liver when evaluated against arthritic animals (Group 2). No remarkable change was observed in the erythrocytes GSH content in indomethacin and VNE treated group (Group 3 and 4) as compared to control (Group 1) and arthritic group (Group 2). 3.11. Determination of carbonyl content Protein carbonyl content was significantly (p< 0.001) higher in liver and plasma of arthritic (Group 2, Table 5) and VNE treated animals (Group 4) when compared to control (Group 1). However, the decline level of carbonyls was observed with indomethacin in liver (p< 0.001), whereas the change in carbonyl content was insignificant in liver and plasma of the VNE group (Group 4) as compared to arthritic group (Group 2).
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