The Effects Of Naringenin And Alpinetin In Nrf2/Phase Ⅱ Detoxifying Enzymes Activities And Studies On Atherosclerotic Lesions

Objective:The study aims to investigate the effects of naringenin and alpinetin in nuclear factor erythroid 2-related factor 2(Nrf2)/phase Ⅱ detoxifying enzymes activities and evaluate their role on atherosclerotic lesions,providing more effective targeted therapy drugs for the treatment of atherosclerosis.Methods:Western blot was used to detect the expression of Nrf2 in the cytoplasm and nucleus,and the expression of related phase Ⅱ detoxification enzymes and inflammatory factors in cells and livers.Vascular immunofluorescence and reverse transcription-qPCR were used to detect the expression of related inflammatory factors in the aortic root.The activities of phase Ⅱ detoxifying enzymes were measured by commercial kits.Co-immunoprecipitation technology was used to detect the interaction between Nrf2 and kelch-like ECH-associated protein 1(Keap-1).Confocal microscope was used to detect the position of Nrf2.Oil red O and HE staining of blood vessels and aortic roots were used to analyze the degree of atherosclerosis.Results:Narigenin and alpinetin promoted the dislocation of Nrf2 from Keap1 compound and increased Nrf2 nuclear accumulation in RAW264.7 macrophages.Naringenin and alpinetin up-regulated expression of phase Ⅱ detoxifying enzymes such as NAD(P)H quinone oxidoreductase 1(NQO1),glutathione S-transferase(GST)and glutamate-cysteine ligase(GCL).Narigenin also reduced levels of cytokines in macrophages.Moreover,the Nrf2 inhibitor ML385 reduced phase Ⅱ detoxifying enzymes expression and increased cytokine levels.In addition,we found naringenin increased the expression and activities of liver phase Ⅱ detoxifying enzymes(NQO1,GST and GCL)and reduced aortic inflammation in atherosclerotic model mice.The effects were dependent on Nrf2 activity.Alpinetin inhibited lipid absorption and the formation of foam cells through Nrf2 in macrophages.Meanwhile,alpinetin increased the activities of NQO1,GST and GCL in the livers of atherosclerotic model mice by Nrf2,and reduced lipid accumulation and plaque lesions in aortic.Conclusions:Naringenin activates Nrf2 and promotes phase Ⅱ detoxifying enzymes activities,which leads to inhibition of vascular inflammation.Alpinetin inhibits lipid absorption and foam cell formation by activating Nrf2,promotes the expression of phase Ⅱ detoxification enzymes and reduces lipid accumulation and plaque lesions in aortic.