去天冬氨酸血管紧张素-1对人脐静脉内皮细胞的保护作用

Protective effects of des-aspartate-angiotensin-1 on endothelial cells of human umbilical vein

目的观察天冬氨酸血管紧张素-1(des-aspartate-angiotensin1,DAA-1)对过氧化氢致人脐静脉内皮细胞损伤的保护作用。方法实验分为正常对照组,过氧化氢模型组,过氧化氢加DAA-1组(低剂量、中剂量、高剂量组)。倒置显微镜下观察细胞形态、密度。噻唑蓝(MTT)法检测细胞活力,比色法测定细胞培养液中超氧化物歧化酶,活性、丙二醛及乳酸脱氢酶浓度。免疫细胞化学方法测定组织纤维蛋白溶酶原激活物,Ⅰ型纤溶酶原激活物抑制因子表达量。逆转录酶-聚合酶链反应(RT-PCR)方法测定细胞内皮素1和血栓调节蛋白mRNA的表达。结果DAA-1能明显抑制过氧化氢所致的细胞活力的降低[0.443±0.024vs.0.317±0.024,P<0.01],减少细胞内乳酸脱氢酶释放[(102.35±9.87)U/Lvs.(242.78±16.10)U/L,P<0.01],提高超氧化物歧化酶活力[(19.10±2.33)U/mLvs.(8.16±3.48)U/mL,P<0.01],并呈剂量依赖性作用;能明显减少内皮细胞在氧化过程中丙二醛的生成[(1.35±0.19)U/mLvs.(2.33±0.48)U/mL,P<0.01],增加组织纤维蛋白溶酶原激活物蛋白表达,减少Ⅰ型纤溶酶原激活物抑制因子蛋白表达;并能明显抑制内皮素mRNA的表达和提高血栓调节蛋白mRNA的表达。结论DAA-1对过氧化氢所致人脐静脉内皮细胞损伤具有明显的保护作用。

Objectives To evaluate the protective effects of des-aspartate-angiotensin-1 （DAA-1） on human umbilical vein endothelial cells （ECV-304） injured by hydrogen peroxide in vitro. Methods The activity of ECV-304 was monitored by MTT assay. The activity of superoxide dismutase （SOD） level in culture medium was measured by colorimetric,so was malondialdehyde （MDA） and lactic dehydrogenase （LDH） content of ECV-304. The expression of tissue plasminogen activator and plasminogen activator inhibitor-1 was monitored by the way of immuocytochemistry.The expression of thrombomoduline and endothelins-1 was measured by means of reverse transcription polymerase chain reaction. Results DAA-1 inhibited the activity of ECV-304 injured by hydrogen peroxide [0.443±0.024 vs. 0.317± 0.024,P〈0.01],reduced MDA[（1.35±0.19）U / mL vs. （2.33±0.48）U / mL,P〈0.01] and LDH[（102.35±9.87）U / L vs. （242.78±16.10）U / L,P〈0.01] content and enhanced the activity of SOD [（19.10±2.33）U / mL vs. （8.16±3.48）U / mL,P〈0.01]. They could also reduced the expression of plasminogen activator inhibitor-1 and increase the expression of tissue plasminogen activator.Furthermore,DAA-1 reduced the expression of endothelins-1 and increase the expression of thrombomoduline. Conclusions DAA-1 has obviously protective effects on ECV-304 injured by hydrogen peroxide.