Compared to the vehicle treated group (V group), a significant increase in the percentage of positive cells/the percentage intensity of the total apoptotic nuclei was observed following 24 h of single dose of B(a)P [subgroup BP(+24h)] in liver whereas in the lungs, it was similar to the vehicle treated group (Figs. 4A and 4B). It was observed that compared to subgroup BP(+24h), mice on control diet for 24, 72 and 120 h [subgroups
BP(+48h), BP(+96h), BP(+144h)] showed an increase in apoptotic cells as judged by the percentage of TUNEL positive apoptotic cells (apoptotic index) and/or the percentage intensity of total apoptotic nuclei in the liver and lungs of mice. Interestingly, mice that were shifted to the 0.05% curcumin diet and killed at 72 and 120 h [subgroups
BP(+96h) + C 72 h, Everolimus cost BP(+144h) + C 120 h] showed further increase in B(a)P-mediated apoptosis as seen by an increase in numbers of apoptotic cells as well as the percentage intensity of total apoptotic nuclei compared to BP(+24h) and respective Sirolimus concentration time-matched controls [subgroups BP(+96h) and BP(+144h)] (Figure 4 and Figure 5). These observations thus suggest that dietary curcumin further enhances the B(a)P-induced apoptosis, which would indirectly confer protection due to increased removal of adduct containing cells. As observed in experiment 1, 5-10% and 20-35% of total apoptotic cells (apoptotic index) were detected in the liver and lung tissues of vehicle [V(+24h), V(+8d), V(+15d), V(+29d)] or vehicle + curcumin [V(+8d) + C 7d, V(+15d) + C 14d, V(+29d) + C 28d]-treated subgroup, respectively 4-Aminobutyrate aminotransferase (Figs. 4 C and 4D). It was observed that compared to subgroup BP(+24h), mice on the control diet for 7 days [subgroup BP(+8d)] showed an increase in apoptosis as judged by an increased percentage of positive cells (apoptotic index) and/or the percentage intensity of
total apoptotic nuclei in the liver and lungs of mice whereas a relative decrease in apoptosis in the liver was observed in mice on the control diet for 14 and 28 days [subgroups BP(+15d), BP(+29d)] (Figure 4 and Figure 5). Interestingly, mice that were shifted to the 0.05% curcumin diet and killed at 7, 14 and 28 days did not show significant difference in the level of apoptosis in the liver and lungs of mice compared to BP(+24h) and respective time-matched controls [subgroups BP(+8d), BP(+15d), BP(+29d)]. However at 8 days, in the liver mice showed a decrease in the percentage of positive apoptotic cells (apoptotic index) and/or the percentage intensity of total apoptotic nuclei (Figure 4 and Figure 5). An observed decrease in DNA adducts without enhancement in the levels of apoptosis in the liver and lungs suggests a role of DNA repair and/or dilution of BPDE-DNA adducts in tissue cells. Further, to confirm and compliment the post-treatment effects of dietary curcumin in enhancement of apoptosis measured by TUNEL assay, protein levels of apoptosis-related markers were analyzed in the liver and lungs of mice by immunoblotting.