Figure 2.

Curcumin reduces αS-induced intracellular ROS generation and inhibits caspase-3 activation in SH-SY5Y cells. SH-SY5Y cells were incubated with Tris buffer, oligomeric αS, αS+curcumin and curcumin and the intracellular ROS and caspase-3 activity were determined using cell based assays. (A) Intracellular ROS was determined by DCF fluorescence. DCFH-DA was then added to each well and the plate was incubated at 37°C for an additional 1 hr. The fluorescence intensity of DCF was measured at Ex485 nm and Em535 nm, respectively. The increase in DCF fluorescence was expressed as a percentage of the control and is a direct measurement of intracellular ROS due to the oxidation of DCFH-DA to DCF by intracellular ROS. (B) Caspase-3 activity was determined by the absorbance of pNA substrate. After 24 h of treatment, the cells were detached, lysed and an equal protein loading was added to the 2× reaction buffer with DTT and DEVD-pNA substrate. After 1 hr of incubation at 37°C, the absorbance intensity was measured at 405 nm and the caspase-3 activity was reported percentage of the Tris buffer control. Data was analyzed using one way ANOVA followed by Bonferroni post-hoc test and reported as mean ± SE, n = 4. **p < 0.01, ***p < 0.001, compared with the untreated samples; and ##p < 0.01, ##p < 0.001, compared with αS-treated samples

Wang et al. BMC Neuroscience 2010 11:57   doi:10.1186/1471-2202-11-57
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