Caprnda Martin, Kubatka Peter, Saxena Sandeep, Valaskova Jela, Stefanickova Jana, Kobyliak Nazarii, Zulli Anthony, Kruzliak Peter
First Department of Internal Medicine, Faculty of Medicine, Comenius University, Bratislava, Slovakia
Department of Medical Biology, Jessenius Faculty of Medicine, Comenius University in Bratislava, Martin, Slovak Republic
Folia Med (Plovdiv). 2017 Jun 1;59(2):183-189. doi: 10.1515/folmed-2017-0029.
Diabetic retinopathy is a serious sight-threatening complication which is manifested by excessive angiogenesis in diabetic patients.
We hypothesize that cultured Rhesus monkey retinal endothelial cells (RhRECs) respond to high glucose with a change in cell proliferation and vascular endothelial growth factor (VEGF) secretion.
In our study, 20 000 cells per well were treated without glucose or with 5.5 mM, 18.5 mM and 30 mM glucose for 24 hours. Viable cells were counted using trypan blue dye exclusion method. VEGF concentrations were measured in cell media by ELISA method.
The number of viable cells incubated with 5.5 mM glucose increased significantly by 53.7% after 24 hours. In comparison, the number of viable cells decreased by 2.8% at 18.5 mM of glucose and by 20.4% at 30 mM of glucose after 24 hours of incubation. In contrast to this effect of glucose on the number of viable cells, a significant increase in VEGF levels (pg/mL) in the cell media with a glucose concentration of 0 mM compared to 5.5 mM of glucose was found. VEGF secretion in cell medium with 18.5 and 30 mM of glucose increased non-significantly in comparison with euglycemic levels.
Our results show that viability of retinal endothelial cells and VEGF release are highly responsive to changes in glucose concentration. Such glucose-induced changes in retinal endothelial cells may negatively impact the integrity of the microvasculature in the diabetic retina leading to angiogenesis and microaneursym.
糖尿病视网膜病变是一种严重的致盲并发症,表现为糖尿病患者血管生成过多。
我们假设培养的恒河猴视网膜内皮细胞(RhRECs)对高糖的反应是细胞增殖和血管内皮生长因子(VEGF)分泌发生变化。
在我们的研究中,每孔接种20000个细胞,分别用无糖培养基或含5.5 mM、18.5 mM和30 mM葡萄糖的培养基处理24小时。使用台盼蓝染料排斥法对活细胞进行计数。通过ELISA法测定细胞培养基中的VEGF浓度。
用5.5 mM葡萄糖培养24小时后,活细胞数量显著增加了53.7%。相比之下,在18.5 mM葡萄糖浓度下培养24小时后,活细胞数量减少了2.8%,在30 mM葡萄糖浓度下减少了20.4%。与葡萄糖对活细胞数量的这种影响相反,发现与5.5 mM葡萄糖相比,葡萄糖浓度为0 mM时细胞培养基中VEGF水平(pg/mL)显著升高。与正常血糖水平相比,含18.5 mM和30 mM葡萄糖的细胞培养基中VEGF分泌无显著增加。
我们的结果表明,视网膜内皮细胞的活力和VEGF释放对葡萄糖浓度的变化高度敏感。视网膜内皮细胞中这种由葡萄糖诱导的变化可能会对糖尿病视网膜微血管的完整性产生负面影响,导致血管生成和微动脉瘤形成。