Figure 3

Effects of FA on LPS-induced BMEC ROS generation and changes in the MMP and Bax/Bcl-2 expression. Cells were challenged with LPS (50 μg/mL) for 12 h after an incubation with the indicated concentrations (0, 5, 10, or 15 μg/mL) of FA or serum-free media for 2 h (A–D). A The intracellular ROS level was measured using flow cytometry. One plot from three independent experiments is shown, and ROS levels were measured. The data are presented as means ± SEM (n = 3). ^###P < 0.001 compared with the control group, *P < 0.05 compared with LPS group, and **P < 0.01 compared with the LPS group. B The cellular MMP was determined by performing a bivariate JC-1 analysis using flow cytometry. Signals in the upper right quadrant represent the aggregated red-emitting JC-1 in the matrix of undamaged mitochondria with a normal MMP, while signals in the lower right quadrant represent the green-emitting JC-1 monomers in the cytoplasm of cells with damaged mitochondria. C The ratios of red to green fluorescence intensity in different groups are presented, and the values represent the magnitude of the MMP. The data are presented as means ± SEM (n = 3). ^###P < 0.001 compared with the control group, *P < 0.05 compared with the LPS group, and ***P < 0.001 compared with the LPS group. D The Bax/Bcl-2 expression levels were quantified using Western blot analysis. The data are presented as means ± SEM (n = 3). ^###P < 0.001 compared with the control group, *P < 0.05 compared with the LPS group, and ***P < 0.001 compared with the LPS group. E The dynamic changes in cellular MMP and intracellular ROS levels were analysed using flow cytometry after stimulation with LPS (50 μg/mL) for different times. The data are presented as means ± SEM (n = 3). *P < 0.05 compared with the control group, **P < 0.01 compared with the control group, and *** P < 0.001 compared with the control group.