Fig. 5

1,25(OH)₂D₃ alleviated endothelial inflammation in high glucose and PM-treated HUVECs. HUVECs were pretreated with high glucose (30 mM) for 24 h and then treated with PM (50 μg/ml) for 8 h. A, B The expression of ICAM-1 and VCAM-1 was detected using Western blotting. C The levels of IL-6 were measured using ELISA. d Representative fluorescence images showing the adhesion of fluorescein-labeled THP-1 cells to HUVECs. E, F HUVECs were pretreated with 100 nM 1,25(OH)₂D₃, 500 nM MitoTEMPO and 10 mM NAC for 12 h and 1 h, respectively. The expression of ICAM-1 and VCAM-1 was detected using Western blotting. G HUVECs were pretreated with 100 nM 1,25(OH)₂D₃, 500 nM MitoTEMPO and 10 mM NAC for 12 h and 1 h, respectively. Representative fluorescence images showing fluorescein-labeled THP-1 cells adhering to HUVECs. H, I HUVECs were pretreated with 100 nM BAF or 10 μM CQ for 1 h before PM exposure. The expression of ICAM-1 and VCAM-1 was detected using Western blotting. J HUVECs were pretreated with 100 nM BAF or 10 μM CQ for 1 h before PM exposure. Representative fluorescence images showing fluorescein-labeled THP-1 cells adhering to HUVECs. K Autophagy flux analysis with tandem sensor RFP-GFP-LC3B. HUVECs received the aforementioned treatment. Cells were stained with DAPI. The autophagosomes and autolysosomes were shown as the yellow dots and red dots, respectively. Bar = 25 μm. The number of autophagosomes and autolysosomes were counted per cell. L HUVECs were transfected with a scrambled siRNA (siScr) or siRNA targeting Bnip3 (siBnip3) and then exposed to high glucose and PM. AO was used to detect the formation of autolysosomes. Bar = 50 μm. M HUVECs were transfected with a scrambled siRNA (siScr) or siRNA targeting Bnip3 (siBnip3), followed by high glucose and PM exposure. The expression of ICAM-1, VCAM-1 and Bnip3 was detected using Western blotting. N HUVECs were transfected with a scrambled siRNA (siScr) or siRNA targeting Bnip3 (siBnip3), and then exposed to high glucose and PM. Representative fluorescence images showing the adhesion of fluorescein-labeled THP-1 cells to HUVECs. *P < 0.05 compared with the control group; ^†P < 0.05 compared with the HG group or HG + PM group; ^#P < 0.05 compared with the PM group; ^§P < 0.05 compared with the HG + PM group or HG + PM + siScr group