Figure 1.
MGO concentration-dependently induced cell injury in the cultured HBMEC.
(A) HBMEC were incubated with different concentration of MGO (100 µmol/l, 500 µmol/l, 1 mmol/l, 2 mmol/l and 5 mmol/l) for 24 h. (B) HBMEC were incubated with MGO (2 mmol/l) for 3, 6, 12, 24 h. Cell viability was determined by MTT assay. *p<0.05 versus control group. n = 3 repeats.
Figure 2.
Edaravone protected MGO-induced cell injury in the cultured HBMEC.
(A) HBMEC were incubated with edaravone (10 µmol/l, 50 µmol/l, 100 µmol/l), aminoguanidine (1 mmol/l) for 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, cell viability was determined by MTT assay. (B) Cell counts were assessed by trypan blue exclusion. (C) HBMEC were incubated with edaravone (100 µmol/l), aminoguanidine (1 mmol/l) 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, cell apoptosis was detected by rhodamine 123 staining and flow cytometer. *p<0.05 versus control group. #p<0.05 versus MGO group. n = 3 repeats. AG represented aminoguanidine. ED represented edaravone.
Figure 3.
Edaravone inhibited MGO-induced AGE-RAGE axis and cellular oxidative stress.
(A) BSA-MGO assay. (B) The modulation of MGO-mediated BSA modification as revealed by SDS-PAGE. Results are means ± S.D. (C, D, E) HBMEC were incubated with edaravone (100 µmol/l), aminoguanidine (1 mmol/l) 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, RAGE, AGEs were determined by Western blotting. (F) HBMEC were incubated with edaravone (100 µmol/l), aminoguanidine (1 mmol/l) 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, cellular oxidative stress was determined by ROS release. *p<0.05 versus control group. #p<0.05 versus MGO group. n = 3 repeats. AG represented aminoguanidine. ED represented edaravone.
Figure 4.
Edaravone protected MGO enhanced OGD-induced injury in the cultured HBMEC.
(A) HBMEC were incubated with MGO (2 mmol/l) for 24 h before 3 h OGD. Cell viability was determined by MTT assay. (B) HBMEC were incubated with edaravone (100 µmol/l), aminoguanidine (1 mmol/l) 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, the cultured HBMEC was incubated under OGD condition for another 3 h. Cell viability was determined by MTT assay. (C) Cell mortality was determined by LDH assay. (D, E, F) HBMEC were incubated with edaravone (100 µmol/l) 20 min before MGO (2 mmol/l) treatment. After 24 h MGO treatment, the cultured HBMEC was incubated under OGD condition for another 3 h. RAGE, AGEs were determined by Western blotting. $p<0.05 versus MGO group. *p<0.05 versus control group. #p<0.05 versus OGD group. &p<0.05 versus MGO+OGD group. n = 3 repeats. AG represented aminoguanidine. ED represented edaravone.