Figure 3.

ALCAR+LA supplementation lowers 2,′7′-dichlorofluorescin (DCF) appearance. Young and old rats were treated with or without ALCAR+LA, and hepatocytes were isolated as described. Cells (40,000) were incubated with dichlorofluorescin diacetate (20 μM, final concentration), and the rate of fluorescence appearance was measured by using a Cytofluor fluorescent plate reader (as described in Materials and Methods). Because of the difference in oxygen consumption characteristics, all rates of fluorescence were expressed in relation to oxygen consumption. Results show that ALCAR+LA supplementation significantly lowered the age-related increase in DCF appearance, indicating lower ROS and/or reactive nitrogen species in hepatocytes under this feeding regimen. Columns denoted by the same letter are statistically different (P ≤ 0.05) from each other.