高糖环境对体外培养糖尿病大鼠大血管内皮细胞产生转化生长因子-β的影响

Effect of High Glucose Condition on the Secretion of TGFβ by Culture d Macrovascular Endothelial Cells of the Diabetic Rat

目的 探讨体外高糖环境对糖尿病大鼠大血管内皮细胞及内皮细胞株产生转化生长因子 - β( TGF- β)的影响。方法 1制备糖尿病大鼠动物模型 ;2培养糖尿病大鼠大血管内皮细胞 ;3分别用含不同浓度葡萄糖 ( 12 .5 m mol/ L、2 5 m mol/ L、5 0 mm ol/ L D-葡萄糖 )、高糖加肿瘤坏死因子 - α( TNF- α)和高糖加胰岛素的 10 %小牛血清 RPMI16 40液培养糖尿病大鼠主动脉血管内皮细胞 48小时 ,取上述培养液分别用生物学方法测其 TGF- β1水平 ,采用放射免疫法测其内皮素 ( ET)的水平。结果 高糖环境下血管内皮细胞分泌的总 TGF- β1 ( 40 6± 2 9pg/ml)及活性 TGF- β1 ( 16 1± 2 4pg/ m l)含量与对照组 (总 TGF- β1 189± 2 3pb/ m l,活性 TGF- β1 39.5± 8pg/ m l)相比明显增加 ( P均 <0 .0 5 )。TNF- α可以加强高糖的这种刺激作用 ,而胰岛素处理可以抑制这种作用。高糖刺激后 ,培养上清中 ET含量明显增加 ( P<0 .0 5 ) ,TNF- α可以加强这种作用 ,胰岛素却能抑制这种作用。结论 TGF-

Objective This study was designed to examine the secretion of TGF β by cultured rat thoracic aortic endothelial cells of the diabetic rats under high gluocse condition. Methods First we created diabetic rat models, then the thoracic aortic endothelial cells of th e diabetic rats were isolated and cultured in 10% FBS RPMI 1640 media with vario us high glucose concentrations (12.5mmol/L,25mmol/L or 50mmol/L D glucose), h i gh glucose and TNF α or high glucose and insulin for 48 hours, and the supern a tant obtained from the cultured aoritc endothelial cells was collected. The leve l of TGF β 1 in the supernatant was measured using a standard bioassay,which utilizes the TGF β 1 sensitive Mv1Lu(CLL 64) Mink lung epithelial cell li ne, and the produ ction of endothelin (ET) in the supernatant was also assessed by RIA (radio immo l/Lunoassay). Results Both total TGF β 1 (406±29 vs 189±23 pg/ ml, P <0.05) and active TGF β 1 (161±24 vs 39.5±8pg/ml) secreted by the cultured aortic endothelial cells under high glucose condition (25mmol/L) were significa ntly increased as compared with control. TNF α could enhance the effect of hi gh glucose on the secretion of TGF β 1, whereas insulin could inhibit it. Wit h various high glucose concentrations, the levels of ET were increased. Also TNF αenhanced it and in sulin inhibited it. Conclusion TGF β 1 may be involved in the development of diabetic macrovascular complication.