Inhibitory Effect of Paeonol on Apoptosis, Oxidative Stress, and Inflammatory Response in Human Umbilical Vein Endothelial Cells Induced by High Glucose and Palmitic Acid Induced Through Regulating SIRT1/FOXO3a/NF-κB Pathway.

ABSTRACT

Reactive oxygen species (ROS) induced by high glucose and high fat of diabetes mellitus (DM) finally caused the occurrence and progression of atherosclerosis and other macrovascular complications. Paeonol (Pae) exhibits anti-inflammation, antioxidation, and antiatherosclerosis activities. However, the role of Pae in diabetic cardiopathy has not been fully understood. Therefore, we aimed to investigate the role of Pae in diabetic cardiovascular diseases. Human umbilical vein endothelial cells (HUVECs) were exposed to high glucose and palmitic acid (HG/HP), a model DM environment and different doses of Pae. The viability and apoptotic rate of HUVECs were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry assay, respectively. Oxidative indicators (ROS, malondiadehyde [MDA], superoxide dismutase [SOD]), and inflammatory cytokines (tumor necrosis factor-α, interleukin-1ß, and interleukin-6) were detected by 2,7-dichlorodihydrofluorescein diacetate, colorimetry, and enzyme-linked immunosorbent assay. The protein levels of Sirtuin type 1 (SIRT1), Bcl-2, Bax, Cleaved caspase-3, p-p65, and p-65 were detected by Western blot. The mRNA levels of Bcl-2 and Bax were detected by quantitative real-time polymerase chain reaction. The acetylation and protein levels of forkhead box O3a (FOXO3a) were detected by immunoprecipitation assay. SIRT1 silencing was used to confirm the role of Pae in the resistance to apoptosis, oxidative stress, and inflammatory response. Pae increased SIRT1 expression, cell viability, and SOD activity and suppressed apoptosis, the levels of p-p65/p-65, ROS, MDA, and inflammatory cytokines, and the expression of acetylated-FOXO3a induced by HG/HP in HUVECs. SIRT1 silencing abrogated the effect of Pae on HG/HP-mediated HUVECs. Inhibitory effect of Pae on apoptosis, oxidative stress, and inflammatory response in HUVECs induced by HG/HP induced through regulating SIRT1/FOXO3a/NF-κB pathway.