摘要
目的:研究维生素B1(thiamine)通过改变糖酵解和细胞复制对高浓度葡萄糖所致体外培养的牛视网膜微血管内皮细胞(BREC)损伤的保护作用,探讨维生素B1的保护作用。方法:取2～3代体外培养的牛视网膜微血管内皮细胞,将培养板上的细胞随机分为4组,DMEM培养基含不同葡萄糖浓度(5.5,20.0mmol/L),同时分别加与不加维生素B1(150μmol)。7d后检测细胞活力和上层培养液中的LDH的含量;3H-TdR掺入法测定DNA合成以及流式细胞仪测定牛视网膜微血管内皮细胞细胞周期,观察高糖对牛视网膜微血管内皮细胞的损伤作用并观察维生素B1对其的保护作用。结果:与正常组(葡萄糖5.5mmol/L)相比,高糖组(葡萄糖20.0mmol/L)的细胞活力明显降低,培养液中的LDH含量显著增加,细胞DNA合成减少,细胞凋亡率增加。而加入维生素B1(150μmol)能够显著降低细胞的损伤程度。结论:高浓度葡萄糖对牛视网膜微血管内皮细胞有显著的损伤作用,维生素B1能减轻高糖对内皮细胞的损伤,其保护作用可能是通过改变糖酵解和细胞复制来实现的。
AIM: To evaluate effect of thiamine on the damage of high glucose on bovine retinal endothelial cells (BREC) in vitro so as to study the protective function of thiamine.· METHODS: Endothelial cells obtained from bovine retinas were cultured in vitro. The effect of different glucose concentrations (5.5, 20mmol/L) with or without thiamine (150μmol) on BREC was evaluated by measurement of MTT and lactate dehydrogenase (LDH) released into the culture medium and 3H-thymidine incorporation after 7d respectively. The phases of the cells cycle were analyzed by flow cytophotometry under these conditions.· RESULTS: High glucose had an effective inhibition rate on endothelial cells. When thiamine was added to cells kept under high glucose conditions, the number of surviving cells was significantly increased; the reduced cell proliferation and 3H-thymidine incorporation appeared to be corrected. LDH leakage confirmed the increased production of this metabolite by endothelial cells in high glucose, which was reduced by thiamine. The endothelial replication was corrected by thiamine.· CONCLUSION: High glucose has a significant inhibitive effect on bovine retinal endothelial cells. The beneficial effect of thiamine limits this damage. Thiamine corrects abnormal levels of glycolytic metabolites and delays endothelial replication. ·
出处
《国际眼科杂志》
CAS
2005年第1期77-79,共3页
International Eye Science
