摘要
目的:研究高浓度葡萄糖对体外培养的视网膜色素上皮(retinal pigment epithelium,RPE)细胞增生以及活性氧(reactive oxygen species,ROS)生成的影响。方法:体外培养人RPE(ARPE-19),随机分成4组,即正常对照组用含5.6mmol/L葡萄糖的DMEM/F12培养液,高糖组用30mmol/L葡萄糖的DMEM/F12培养液,高糖+SB203580组用p38-MAPK特异性阻断剂SB20358010μmol/L预处理30min后,再用含30mmol/L葡萄糖的DMEM/F12培养液,甘露醇组(渗透压对照组)用含5.6mmol/L葡萄糖和24.4mmol/L甘露醇的DMEM/F12培养液。各组培养48h用MTT法检测各组视网膜色素上皮细胞的增生活力,用CM-H2DCFDA荧光染色检测RPE细胞中ROS的产生量。结果:与对照组相比,高糖培养人视网膜色素上皮细胞48h可以导致RPE细胞的损伤,抑制RPE细胞增殖,并使ROS生成增加,在一定程度上,与p38-MAPK途径的激活有关。结论:高糖培养ARPE-19可致细胞损伤,抑制增殖,并使ROS生成增加。
AIM : To investigate the effects of high glucose condition on the proliferation and reactive oxygen species (ROS) expression in cultured human retinal pigment epithelial (RPE) cells in vitro. METHODS: Human RPE cells were cultured in vitro and divided into four groups: control group with DMEM/F12 culture solution including 5. 6mmol/L glucose; high glucose group with DMEM/F12 culture solution including 30mmol/L glucose; high glucose plus SB203580 group with DMEM/F12 culture solution including 30mmol/L glucose plus 10μmol/L SB203580 the specific in hibitor of p38 mitogen activated protein kinase ( p38-MAPK ); mannitol group with DMEM/F12 culture solution including 5.6mmol/L glucose and 24. 4mmol/L mannitol. Cell viability was assessed by the MTT assay. ROS change in RPE cells in response to high glucose was detected with flow cytometry. RESULTS: Compared to controls , the treatment of RPE cells with 30mmol/L glucose caused an obviously decrease of cellar viability. After pretreated with SB203580, cell viability was elevated compared with high glucose group. The ROS increased in high glucose group. CONCLUSION: High glucose could damage RPE cells . The mechanisms may be that high glucose could evoke ROS expression via p38-MAPK pathway.
出处
《国际眼科杂志》
CAS
2010年第3期453-455,共3页
International Eye Science