高糖对人脐静脉内皮细胞表面血栓调节蛋白表达及活性的影响

Effect of High Glucose on the Expression and Activity of Thrombomodulin in Human Umbilical Vein Endothelial Cells

目的观察高糖对血栓调节蛋白在原代人脐静脉内皮细胞的表达和活性的影响。方法体外培养人脐静脉内皮细胞,实验分为三组:1对照组;210 mmol/L葡萄糖组;320 mmol/L葡萄糖组,孵育细胞24 h。分别用流式细胞术、实时荧光定量聚合酶链反应和酶标仪检测血栓调节蛋白的蛋白、m RNA表达及活性强度。结果20 mmol/L葡萄糖组和10 mmol/L葡萄糖组相比于对照组在血栓调节蛋白的蛋白(41.38±3.41、32.60±2.59比27.96±1.58)、m RNA(2.05±0.19、1.44±0.32比1)水平上均升高(P<0.05),20 mmol/L葡萄糖组与对照组相比在血栓调节蛋白活性上也增强(0.4157±0.0129比0.3957±0.0100,P<0.05),但10 mmol/L葡萄糖组与对照组在血栓调节蛋白活性上差异无显著性。结论高糖可增加血栓调节蛋白表达,并增强其活性,提示这可能为内皮细胞对于高糖的一种防御机制。

Aim To observe the effect of high glucose on the expression and activity of thrombomodulin in human umbilical vein endothelial cells( HUVEC). Methods HUVEC were incubated in vitro,sub-experiment: 1control group; 210 mmol / L glucose group; 320 mmol / L glucose group,all incubated for 24 h. Flow cytometric and RTPCR technique were used for detecting thrombomodulin protein and m RNA,respectively. Microplate Reader was used for detecting thrombomodulin activity. Results Compared with control group,both m RNA( 2. 05 ± 0. 19、1. 44 ± 0. 32 vs1) and protein( 41. 38 ± 3. 41、32. 60 ± 2. 59 vs 27. 96 ± 1. 58) levels of thrombomodulin increased in 20 mmol / L glucose group and 10 mmol / L glucose group( P < 0. 05) and the thrombomodulin activity enhanced( 0. 4157 ± 0. 0129 vs 0. 3957± 0. 0100,P < 0. 05) in HUVEC of 20 mmol / L glucose group. Conclusion Exposure to high glucose increased the expression of thrombomodulin and enhanced the thrombomodulin activity in HUVEC. The upregulation of thrombomodulin in HUVEC cultivated in high glucose conditions may represent a defense mechanism against the stress.