Fisetin Alleviates Neointimal Hyperplasia via PPARγ/PON2 Antioxidative Pathway in SHR Rat Artery Injury Model

Figure 3

Fisetin inhibited AngII-induced ROS production in VSMCs through PON2. (a) Total ROS in cells was measured using the fluorescence probe DCFH-DA. Fluorescence intensity was analyzed by Image J. A10 cells were incubated with fisetin (100 μM) or/and AngII (10^-7 M) for 24 hours. (b) A10 cells were incubated with AngII (10^-7 M) for incremental hours (3~24 h). PON2 expression in cells was determined by q-PCR. (c) A10 cells were treated with AngII (10^-7 M)/and fisetin (100 μM) for 24 hours before PON2 mRNA detection by q-PCR. (d) A10 cells were treated with incremental concentrations (0~100 μM) of fisetin and AngII stimulation. Protein expression of PON2 was determined by immunoblotting. (e) A10 cells were transfected with different PON2 siRNAs for 48 hours. PON2 protein expression was determined by Western blotting. siRNA1 was selected for subsequent PON2 interference test. (f) PON2 siRNA-transfected A10 cells were coincubated with AngII and fisetin for 24 hours. Total ROS production was compared with normal PON2 expression cells. Representative images were shown ( vs. control, ^# vs. AngII only, and ^& vs. no siRNA).

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