Fig. 1.
PPARδ activation prevents the radiation-induced increases in intracellular ROS generation, Cox-2 and MCP-1 protein, and IL-1β and TNF-α mRNA levels. A, BV-2 cells were incubated with 10 μM DCFH-DA or 10 μM C369 for 45 min, the probe was washed off using PBS+, and the cells were pretreated with 5 μM L-165041, NAC, or vehicle control for 3 h. The cells were then irradiated with a single dose of 10 Gy of 137Cs γ rays or sham irradiated, and intracellular ROS generation was measured 1 h post-irradiation as described in the Materials and methods. The results are presented as arbitrary fluoresence units. B & C, BV-2 cells were pretreated with 5 μM of L-165401 and irradiated with a single dose of 10 Gy. B, Protein was harvested 7 h post-irradiation and subjected to western blot analysis for Cox-2 or MCP-1; β-actin was used as a loading control. See SFig. 2 for densitometric analysis. C, RNA was harvested 24 h post-irradiation; the expression levels of IL-1β and TNF-α were analyzed using SYBER Green Real-time PCR and normalized with GAPDH expression levels. Changes in gene expression were calculated using the 2ΔΔCt methods (see Materials and methods). Mean ± S.E.M; *, p≤ 0.05 vs. sham-irradiated, #, p≤ 0.05 vs. 10 Gy; n=3.