[Protection of Verapamil against AGE induced apoptosis of human lens epithelial cell]

To investigate the protection of Verapamil against advanced glycation end products (AGE) induced human lens epithelial cells (HLEC) apoptosis.

Experiment study. SRA01/04 (HLEC line) was cultivated and passaged to the third generation and then divided into four groups. A group was named as control group, and B group was named as AGE group (LEC was treated by 20 μmol/L AGE). C group was AGE+SB202190 group (LEC was treated 2 hours by SB2012190 and then treated by 15 μmol/L AGE). D group was AGE+ Verapamil group (LEC was treated 2 hours by 50 μmol/L Verapamil and then treated by AGE). MTT was used to evaluate the cell viability. Flow cytometry with Annexin V-FITC apoptosis detection was used to assess cell apoptosis.The expression of p-p38 and caspase3 was detected by Western blot between groups. One way Chi-square analysis was used for data analysis. LSD-t test was used as comparison between every two groups.

After 24 hours, LEC viability (A570) was (0.28±0.08) in B group, which was significantly lower than A group (0.97±0.05) (LSD-t test, P=0.008). LEC viability in C and D group was (0.79±0.06) and (0.62±0.07) separately, which can partly higher than it was in B group (F=34.52, P=0.001). The apoptosis cells were (19.9±1.1)% in B group, which were significantly higher than they were in A group (2.5±0.6)% (P=0.003). The apoptosis cells in C and D group were (4.23±1.20) and (5.79±1.75) separately, which were significant lower than they were in B group (F=371.61, P<0.01). In additional, expressions of p-p38, Caspase3 proteins in the cells of group B were (223.35±20.15) and (256.77±19.88) separately, which were higher than it were in A group,which were (106.44±10.74) and (100.26±18.65) separately. However, they were (139.17±19.10) and (142.75±23.36) in group C and (154.79±21.87) and (139.79±25.73) in group D (F=248.01, F=76.68; P<0.01), which were lower than they were in A group.

p38 pathway is involved in the apoptotic procedure of LEC induced by AGE. Verapamil can interdict the p38 signal pathway and protect LEC apoptosis. (Chin J Ophthalmol, 2016, 52: 444-448).