Mechanism of Apigenin Improving Vascular Endothelial Cell Dysfunction by High Glucose/High Fat Stress
Objective To investigate the restorative effect of apigenin(AP)on vascular endothelial cell dysfunction induced by high glucose/high fat(HG/HF)stress and its possible mechanism.Methods Human umbilical vein endothelial cells(HUVECs)were divided into 6 groups:the control group received no treatment;group A was treated with HG/HF;group B was treated with HG/HF and 5 μM AP;group C was treated with HG/HF and 25 μM AP;group D was treated with HG/HF and 50 μM AP;group E was treated with HG/HF,25 μM AP,and the PI3K inhibitor LY294002(10 μM).In each group,the effect of wound healing and repair were evaluated using the cell scratch assay;cell proliferation was determined using CCK8;the expression of PI3K-Akt pathway protein was detected by Western blotting;the levels of endothelin-1(ET-1)and NO in cell culture medium were measured using enzyme-linked immunosorbent assay(ELISA)and biochemical assay kits;the reactive oxygen species(ROS)levels were determined by flow cytometry;interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)mRNA levels were detected using qRT-PCR.Results The cell migration rates of groups C and D were significantly higher than that of group A(P＜0.05);the cell proliferation rate of group D was significantly higher than that of group A(P＜0.05);the intracellular ROS levels in groups B,C,and D were all significantly lower than that of group A(P＜0.05).Compared with group A,groups C and D showed significantly reduced ET-1 levels and significantly increased NO levels(both P＜0.05);groups B,C,and D demonstrated significant downregulation of IL-6 mRNA expression levels(all P＜0.05),while groups C and D exhibited significant downregulation of TNF-α mRNA expression levels(P＜0.05);the levels of p-PI3K and p-AKT were significantly elevated in groups C and D(both P＜0.05).The ET-1 level in the cell culture medium of group E was significantly higher than that of group D,while the NO level was significantly lower(both P＜0.05),and the IL-6 and TNF-α mRNA expression levels in group E were significantly upregulated compared to group D(both P＜0.05).Conclusion AP can ameliorate vascular endothelial cell dysfunction induced by HG/HF stress,and this repair effect may be dependent on the activation of PI3K-Akt pathway.
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