Protective Effect of Peroxisome Proliferator-Activated Receptor α Activation against Cardiac Ischemia-Reperfusion Injury Is Related to Upregulation of Uncoupling Protein-3
WY-14643 increased UCP3 expression in H9c2 cells. (a) Treatment of WY-14643 at 50 μM increased expression of PPARα target gene PDK4 mRNA and expression of PPARγ target gene aP2 was not altered after the treatment of WY-14643. (b) Quantitative PCR shows that WY-14643 treatment at 10, 50, and 100 μM for 24 h increased mRNA levels of UCP3 compared with vehicle treatment. versus WY-14643 0 μM (). (c) RT-PCR shows increased UCP3 mRNA expression following WY-14643 treatment. (d) Western blot of UCP3 shows increased protein level after WY-14643 treatment. (e) Treatment of WY-14643 at 50 μM also increased phosphorylation of Akt and STAT3 and protein level of HO-1 as well as UCP3. Representative blots from three independent experiments are shown. Band intensities were quantified using an image analyzer, normalized by expression levels of β-actin, and fold changes are plotted under each band. (f) Transfection of UCP3 siRNA silenced UCP3 mRNA (#2, #4, and #5). (g) Western blot of UCP3 shows that transfection of UCP3 siRNA downregulated UCP3 protein level. #5 was used for further experiment. WY: WY-14643. Rosi: rosiglitazone. HO-1: heme oxygenase-1. SC: scramble.
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