Effect of fisetin on proteome of foam cells / 中草药

ABSTRACT

Objective: To investigate the effect of fisetin (FIS) on foam cells by two-dimensional electrophoresis and mass spectrometry technology, and analyze the molecular mechanism of its inhibition. Methods: MTT method was used to detect the effect of ox-LDL on viability of RAW264.7 cells and fisetin on foam cells separately as well as chemical method was used to detect intracellular cholesterol ester, screening appropriate ox-LDL, and FIS concentration. Oil red O staining displayed accumulation of lipids changed by FIS in the foam cells. Then established proteomic maps of foam cells before and after treatment with FIS by bi-directional electrophoresis, mass spectrometry was adopted to identify differences in the expression of proteins. The expression of Cyt b5 was verified by Western blotting. Results: In our study, 20 μg/mL ox-LDL can successfully induce foam cells, as well as intervention of 100 μg/mL FIS would significantly decrease cholesterol ester and lipid accumulation within the foam cells. Proteomic experiment showed that foam cells treated by FIS had a lower expression of nuclear receptor, calreticulin and transcription elongation factor B, higher levels of glutathione S-transferase, cytochrome b5, Prelamin A/C, NADH dehydrogenase (ubiquinone) 2 flavin protein, lecithin-cholesterol acyltransferase, 78 kDa glucose regulation protein, supervillin, and heat shock protein 60. FIS can significantly improve the expression of Cyt b5 by WB. Conclusion: These data suggest that FIS can significantly inhibit foam cells formation by enhancing the cellular antioxidant and anti-inflammatory capabilities, reducing cellular stress response, as well as decreasing accumulation of intracellular cholesterol, regulation of apoptosis and enhanced immunity to prevent atherosclerosis.