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Effect of fisetin on proteome of foam cells

XU Ying-ling;HU Xiao;FANG Lai;ZHU Ji;WANG Cui;YANG Zhen;LU De-zhao;College of Life Science,Zhejiang Chinese Medical University;Clinical Laboratory,The Third Affiliated Hospital,Zhejiang Chinese Medical University;  
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/m L ox-LDL can successfully induce foam cells, as well as intervention of 100 μg/m L 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 k Da 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.
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