In order to understand the mechanisms of chemopreventive action o

In order to understand the mechanisms of chemopreventive action of saffron, this study used a well-described model of HCC to study the mechanism of the anticancer action of saffron by evaluating its antioxidant, proapoptotic, antiproliferative, and anti-inflammatory effects. 2-AAF, 2-acetyl selleck compound aminofluorene; ABTS, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate); ALT, alanine aminotransferase; ANOVA, analysis

of variance; AST, aspartate aminotransferase; CAT, catalase; COX-2, cyclooxygenase 2; DEN, diethylnitrosamine; DPPH, 1,1-diphenyl-2-picrylhydrazyl; FAH, foci of altered hepatocytes; FRAP, ferric reducing antioxidant power; GGT, gamma glutamyl transpeptidase; GSH, glutathione; GST, glutathione S-transferase; HCC, hepatocellular carcinoma; IL-8, interleukin-8; iNOS, inducible nitric oxide synthase; MDA, malondialdehyde; MPO, myeloperoxidase; NF-κB-p65, nuclear factor-kappa B p-65; P.Carbonyl, protein carbonyl; PI, propidium iodide; p-TNFR1, phosphorylated tumor necrosis factor receptor 1; ROS, reactive oxygen species; SOD, superoxide dismutase; TUNEL, terminal deoxynucleotidyl transferase–mediated dUTP nick-ending labeling. Stigmas of pure saffron were obtained from Golpeech Saffron (register 82558; Mashhad, Iran), and a voucher sample was preserved for reference in the herbarium of United Arab Emirates University. The stigma materials (500 g) were extracted with water or

80% (vol/vol) aqueous ethanol and the mixture was

macerated for 5 days at 4°C. The resulting mixture was then filtered BYL719 solubility dmso and dried under reduced pressure in a rotary evaporator at 40°C to give water and ethanol crude extracts. Photochemical analysis of crocin and safranal derived from saffron was determined PAK5 by high-performance liquid chromatography. The final powder form of saffron extract contained 73.25 mg/g crocin and 33.21 μg/g safranal. The ethanol extract was then administered to animals at a dose of 75, 150, and 300 mg/kg body weight in a volume of 5 mL/kg body weight. The highest dose of saffron used (300 mg/kg) in this study contained 22 mg crocin/kg body weight and 9.96 μg safranal/kg body weight. The experimental hepatocarcinogenesis was initiated by DEN and promoted by 2-AAF, according to the protocol described by Espandiari et al.7 with some modifications. In this model, 96-hours fasting rats were refed as mitotic proliferative stimuli. Then, 24 hours after refeeding, rats were injected intraperitoneally with a single dose of DEN (200 mg/kg body weight) dissolved in saline. Two weeks after DEN treatment, rats received six daily intragastric doses of 2-AAF (30 mg/kg in 1% Tween 80) for promoting hepatocarcinogenesis. All animals received humane care according to the guidelines of the Animal Research Ethics Committee of the UAE University. The experimental design is depicted in Fig.

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