Objective To evaluate antioxidant anti-inflammatory hepatoprotective and vasorelaxant activities of flower buds ethanolic extract. considerable at 10?1 g/L and comparable ((antioxidant and anti-inflammatory activities has been reported. However vasorelaxant anti-inflammatory and hepato-protective activities to the best of our knowledge have never been investigated. In order to better understand the protective effect of buds ethanolic extract on the endothelial and liver functions experiments were performed to determine anti-inflammatory antioxidant and relaxant activities of extract. 2 and methods 2.1 Drugs and chemicals All the reagents and chemicals unless otherwise stated were purchased from Sigma. 2.2 Collection of plant material Fresh flower buds of were collected in March from a remote area in the forest of Tizi Neftah Province of Amizour Department of Bejaia Algeria. The plant was identified by Dr. M.S Benabdelmoumène taxonomist Department of Botany University of Bejaia Algeria. 2.3 Plant sample extraction The fresh flower buds of were air-dried in the shade and ground to a fine powder of 63 μm in diameter. A total of 300 mg of this powder were extracted with ethanol (1: Febuxostat 6 w:v) at room temperature for 24 h. The reunified extractive liquid was evaporated under vacuum. 2.4 Febuxostat Animals Albino mice of either sex weighing around 20 g and purchased from Pasteur Institute (Algiers Algeria) were used in these experiments. They were provided with standard food UTP14C and water buds ethanol extract against aluminum-induced hepatic toxicity was investigated using the modified method of Pan analysis. Differences were considered to be significant at ethanolic extract were measured using the respective standards catechin quercetin and tannic acid to obtain the following equations ethanolic extract were (51.78±4.56) mg catechin Eq/g of extract (13.67±0.34) mg quercetin Eq/g of extract and (228.72±6.90) mg tannic ac Eq/g of extract respectively. 3.2 ABTS assay The results of the ABTS assay Febuxostat indicated that the ethanolic extract of buds of exhibited at a concentration of 100 μg/mL a percentage of (41.05±2.34) in the decolorization of ABTS a moderate activity when compared to the reference quercetin which showed a percentage of (96.5±0.04). 3.3 Acute toxicity Acute toxicity studies did not reveal any toxic symptoms or death in any of the animals at the dose of 200 mg/kg of ethanolic buds extract. 3.4 Anti-inflammatory activity Figure 1 indicates that in the control group (I) the onset of edema [(22.25±4.69)%] occurred as early as 1 h after carrageenan injection and was sustained through the 6 h of observation. On the other hand the extract (200 mg/kg) caused a sharp decrease in paw edema from the 2nd [(21.99±5.58)%] until the 6th hour [(11.14±6.87)%] of Febuxostat treatment (extract. A significant decrease (hepatocytes arranged as radiating plates around the central vein. There was no sinusoidal dilatation or bleeding foci confirming the lack of toxicity of the extract. Figure 2. Photomicrographs of liver sections from mice stained with H&E (×250). On the other hand we observed in both liver sections of mice (Group III) treated with AlCl3 and D-galactose (Figure 2 C1 and C2) which were signs of hepatic damage such as a dilated (green arrow) and congested central hepatic vein (blue arrow) (C1) the presence of some swollen cells increased number of lipid vacuoles (yellow arrow) enlarged nuclei (white arrow) and infiltrating neutrophils (blue arrow) (C2). Most interesting pre-treatment with extract (200 mg/kg) (Group IV) protected almost completely the liver against AlCl3-induced hepatic damage and necrosis as observed in Figure 2(D). Specifically histological examination of the liver of pre-treated animals with plant extract showed that fatty acid changes were less pronounced in comparison with AlCl3-intoxicated mice that have not received extract. 3.6 Effect of P. nigra extract on the levels of eNOS and relaxant action 3.6 Effect on eNOS expression Figure 3 shows that extract did not change the level of phosphorylated eNOS which was the activated form of this enzyme after posttranslational modifications. Figure 3. Western blotting representing the effect of extract on phosphorylated eNOS.