苯磷硫胺对高糖培养的牛视网膜微血管内皮细胞的作用  

Effect of benfotiamine on bovine retinal capillary endothelial cells cultured in high concentration of glucose

作　　者：李斌[1] 刘恒明[1] 曹敏[1] 程铮[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院眼科,武汉430030

出　　处：《眼科研究》2007年第2期124-127,共4页Chinese Ophthalmic Research

摘　　要：目的研究苯磷硫胺对体外高糖环境中培养的牛视网膜毛细血管内皮细胞的生长及细胞凋亡的影响。方法采用MTT法观察不同浓度的苯磷硫胺对高糖培养的牛视网膜毛细血管内皮细胞增生的影响,采用流式细胞分析技术观察选定浓度的苯磷硫胺对高糖环境中培养的牛视网膜毛细血管内皮细胞的凋亡的影响。结果MTT法显示,一定浓度的苯磷硫胺可以降低高糖对牛视网膜毛细血管内皮细胞生长的抑制作用,且呈浓度相关性。在浓度为75μmol/L时OD值差异有统计学意义(P<0.01)。流式细胞分析技术显示,75μmol/L的苯磷硫胺可明显减少牛视网膜毛细血管内皮细胞的凋亡。结论一定浓度的苯磷硫胺对体外高糖环境中培养的牛视网膜毛细血管内皮细胞具有明显的保护作用,其机制可能在于苯磷硫胺能减少细胞凋亡。Objective How to decrease the damage of diabetic retinal capillary is a hot topic in ophthalmology. Benfotiamine is a lipid-soluble derivant of thiamin, and thiamin involves in the critical step of glucose metabolism as a eoferment. The purpose of this paper was to study the effect of benfotiamine on proliferation and apoptosis of bovine retinal capillary endothelial cells cultured in high concentration of glucose. Methods Bovine retinal capillary endothelial cells were cultured in DMEM medium containing 30 mmol/L of glucose and 10% FBS. 0,25,50,75,100μmol/L of benfotiamine were added in above medium. Cultured vascular endothelium ceils were identified by immunochemistry. MTT assay were used to observe the effect of benfotiamine on the cell proliferation. Flow eytometry was used to analyze cell apoptosis during the culture, and Annexin-V marked the earlier apoptosis cells,while PI marked the dead cells. Results The OD value of cultured cells was obviously increased after treated with benfotiamine for 24 hours in over 75μmol/L of concentration groups in comparison with that of the control group （P 〈 0.01 ） ,showing the survival rate of bovine retinal capillary endothelial cells was higher after treated with benfotiamine. No positive correlation was found between the survival rate of bovine retinal capillary endothelial cells and cultured duration （ P 〉 0.05）. Flow eytometry showed that the endothelial cells cultured in high concentration of glucose with 75 μmol/L benfotiamine had lower cell apoptosis rate compared with those cultured in medium with 0,25,50 μmol/L of benfotiamine. Conclusion Benfotiamine can improve the survival rate of bovine retinal capillary endothelial cells cultured in high concentration of glucose and reduce these cell apoptosis in a concentration-dependent manner.

关 键 词：苯磷硫胺 视网膜 毛细血管内皮细胞 凋亡 糖 

分 类 号：R774.1[医药卫生—眼科]