Comparison of the ethanolic and water extracts of saffron showed that the ethanolic extract had the highest phenolic content (67.62 mg gallic acid/g) and the best antioxidant activity. The ascorbic acid equivalent antioxidant capacities of the ethanolic extract were 0.253 and 0.304 mmol/g in the FRAP and ABTS assays,

respectively. Correspondingly, the best free radical scavenging activity, as measured by the DPPH assay, was exerted by ethanolic extract (median inhibitory concentration [IC50] = 2.0 mg/mL) (Table 1). The antioxidant activity of ethanolic saffron extract was also measured in vivo and is summarized in Table 2, which shows the antioxidant status of liver of control and experimental animals. Group 3 (HCC) animals exhibited significant increase Vemurafenib (P < 0.001) in MDA, P.Carbonyl, GSH levels, as well as in MPO and GST activities (P < 0.001) and decrease in CAT (P < 0.01) and SOD (P < 0.001) activities compared to group 1 (control). These findings are consistent with hepatic oxidative stress and damage caused by DEN-2-2AAF. However, as can be seen in Table 2, pretreatment with saffron (groups 4, 5 and 6) significantly attenuated the changes in these oxidative stress markers compared to control. Both medium and high doses of saffron abolished DEN-2-2AAF-induced oxidative stress more effectively than

the lower dose. Surprisingly, GSH levels remained elevated in groups pretreated with saffron and did not return to normal levels as opposed to other oxidative stress markers. Sirolimus in vitro It is well established that GSH synthesis is up-regulated during oxidative damage, inflammation and cell proliferation. It is possible that even in saffron-treated animals, DEN-2-2AAF causes a persistent low level of

oxidative stress as well as low levels of inflammation/proliferation, which in turn causes GSH levels to remain, elevated. Macroscopically, rats in group 3 (HCC) revealed enlarged liver with multiple nodules on the liver surface, the nodule incidence was 75% and the number of nodules per nodule-bearing animal (nodule multiplicity) was 2.33 in this group (Table 3). Groups 4-6, where animals treated with saffron and DEN-2-2AAF, showed a decrease in liver enlargement, nodule incidence and multiplicity. The protective effect of saffron was most dramatic in group 4 rats ZD1839 ic50 (highest dose of saffron), where saffron completely inhibited the appearance of hepatic nodule altogether. Serum activities of GGT, ALT, and AFP were significantly increased in group 3 (HCC) as compared to control rats (group 1), thus indicating liver damage. However, pretreatment with saffron (groups 4-6) caused a significant decrease in the elevation of these proteins (Table 4). Interestingly, higher doses of saffron caused a lesser decrease in the DEN-induced GGT and ALT levels, as opposed to the lowest doses almost reversed these DEN-induced enzyme increase.