Yuan M K, Gao X X, Chang L B
Department of Ophthalmology, Beijing Coal Group General Hospital, Bejing 102300, China.
Zhonghua Yan Ke Za Zhi. 2016 Jun 11;52(6):444-8. doi: 10.3760/cma.j.issn.0412-4081.2016.06.009.
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).
探讨维拉帕米对晚期糖基化终末产物（AGE）诱导的人晶状体上皮细胞（HLEC）凋亡的保护作用。
实验研究。培养SRA01/04（HLEC系）并传代至第三代，然后分为四组。A组为对照组，B组为AGE组（LEC用20μmol/L AGE处理）。C组为AGE+SB202190组（LEC先用SB2012190处理2小时，然后用15μmol/L AGE处理）。D组为AGE+维拉帕米组（LEC先用50μmol/L维拉帕米处理2小时，然后用AGE处理）。采用MTT法评估细胞活力。用Annexin V-FITC凋亡检测流式细胞术评估细胞凋亡。通过蛋白质印迹法检测各组之间p-p38和caspase3的表达。采用单向卡方分析进行数据分析。每两组之间的比较采用LSD-t检验。
24小时后，B组LEC活力（A570）为（0.28±0.08），显著低于A组（0.97±0.05）（LSD-t检验，P=0.008）。C组和D组LEC活力分别为（0.79±0.06）和（0.62±0.07），部分高于B组（F=34.52，P=0.001）。B组凋亡细胞为（19.9±1.1）%，显著高于A组（2.5±0.6）%（P=0.003）。C组和D组凋亡细胞分别为（4.23±1.20）和（5.79±1.75），显著低于B组（F=371.61，P<0.01）。此外，B组细胞中p-p38、Caspase3蛋白表达分别为（223.35±20.15）和（256.77±19.88），高于A组，A组分别为（106.44±10.74）和（100.26±18.65）。然而，C组为（139.17±19.10）和（142.75±23.36），D组为（154.79±21.87）和（139.79±25.73）（F=248.01，F=76.68；P<0.01），低于A组。
p38信号通路参与AGE诱导的LEC凋亡过程。维拉帕米可阻断p38信号通路，保护LEC凋亡。（《中华眼科杂志》，2016，52：444-448）
