吡哆胺对AGEs干预的人RPE细胞的保护作用

Protective effect of pyridoxamine on RPE cells treated with AGEs

摘要:

目的：观察吡哆胺(PM)对高级糖基化终末产物(AGEs)干预的RPE细胞的影响，探讨PM对RPE细胞的保护作用。

方法：取传代至第3代的细胞进行分组：(1)对照组：含100mg/L BSA的DMEM培养基培养48h；(2)AGEs干预组：含200mg/L AGEs的DMEM培养基培养48h；(3)PM组：PM1组：含16mg/L PM+200mg/L AGEs的DMEM培养基培养48h； PM2组：含32mg/L PM+200mg/L AGEs的DMEM培养基培养48h。采用免疫组织化学法检测RAGE蛋白的表达； 荧光染色法观察细胞内ROS的生成； Tunel法检测细胞凋亡情况。

结果：AGEs可明显刺激RPE细胞内RAGE蛋白的表达和ROS的生成，增加细胞凋亡情况，而PM可抑制该过程，且具有一定的浓度依赖性。

结论：PM能够抑制AGEs干预的RPE细胞内RAGE蛋白的表达和ROS生成，减少细胞凋亡，具有一定的细胞保护作用。

Abstract:

AIM: To observe the effects of pyridoxamine(PM)on RAGE, ROS and apoptosis in RPE cells treated with advanced glycation end products(AGEs), and to investigate the protective effect of PM on RPE cells in diabetic retinopathy.

METHODS:Primary cultured human RPE cells, the third generation of cells were synchronized with serum-free Dulbecco-modified Eagle medium for 24h, and then grouped: 1)Control group: cultured with 100mg/L BSA for 48h; 2)AGEs-treated group: cultured with 200mg/L AGEs for 48h; 3)PM group: PM1 group: cultured with 16mg/L PM+200mg/L AGEs for 48h; PM2 group: cultured with 32mg/L PM+200mg/L AGEs for 48h. The expression of RAGE protein was detected by immunohistochemistry. The formation of ROS was observed by fluorescence microscopy. The apoptosis of cells was detected by TUNEL.

RESULTS:The expression of RAGE protein, ROS and apoptosis of RPE cells in PM group were significantly lower than those in AGEs-treated group, and decreased with the increase of PM concentration.

CONCLUSION:Pyridoxamine can inhibit the expression of RAGE and the production of ROS, reduce apoptosis, and have a protective effect on RPE cells.