Impaired Autophagy Induced by oxLDL/β2GPI/anti-β2GPI Complex through PI3K/AKT/mTOR and eNOS Signaling Pathways Contributes to Endothelial Cell Dysfunction

Figure 5

The regulation of PI3K/AKT/mTOR and eNOS signaling pathways in oxLDL/β2GPI/anti-β2GPI complex-mediated endothelial autophagy. HUVECs and bEnd.3 cells were incubated with oxLDL/β2GPI/anti-β2GPI complex for 24 h. LY294002 (10 μM), AZD5363 (1 μM), rapamycin (1 μM), or L-NAME (100 μM) were added 4 h before oxLDL/β2GPI/anti-β2GPI complex treatment. Western blotting analysis of p62 and LC3-II in HUVECs and bEnd.3 cells (a). Quantification of p62 and LC3-II in HUVECs (b) and bEnd.3 cells (c). Representative images (magnification, ×600) of RFP-GFP-LC3 puncta in HUVECs (d) and bEnd.3 cells (e). Yellow puncta (RFP⁺ and GFP⁺) represents autophagosomes, and red puncta (RFP⁺ and GFP^-) represents autolysosomes. Scale bar: 20 μm. , , , and indicate statistically significant differences. ns: nonsignificant differences. All values are denoted as from three independent experiments (), and a representative blot/image was shown.

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