缺氧复氧损伤诱导内皮细胞凋亡及卡托普利的晚期干预作用

目的 探讨缺氧复氧对血管内皮细胞凋亡的影响及卡托普利延迟预处理机制.方法 建立人脐静脉内皮细胞缺氧复氧损伤模型,用流氏细胞仪检测不同的缺氧和复氧时间以及卡托普利晚期预处理,应用缓激肽β2受体阻断剂、PKC阻断剂、一氧化氮合酶阻断剂和NF-κB阻断剂,分别与卡托普利共同孵育细胞,24 h后再对内皮细胞行缺氧复氧损伤,观察不同条件下内皮细胞DNA周期和Annexin V染色率及细胞凋亡情况的变化.结果 单纯缺氧和缺氧复氧均可引起血管内皮细胞凋亡,并且随着单纯缺氧和缺氧后复氧时间的延长,凋亡峰面积和Annexin V染色率逐渐增加.卡托普利晚期预处理后,细胞凋亡明显减少,并且在本实验特定的浓度中（10-2～10-4 mmol/L）,凋亡峰面积随卡托普利剂量的增加而逐渐增强.但给予上面4种阻断剂后,卡托普利的晚期预处理抗凋亡作用均部分消失.结论 单纯缺氧和缺氧复氧均可以导致细胞凋亡,且具有时间依赖性.卡托普利晚期预处理可以减轻内皮细胞凋亡,此作用具有剂量依赖性.这一过程涉及缓激肽β2受体、PKC途径、一氧化氮和核因子的转录等多种因素的参与.

Late protective effect of catopril on vascular endothelial cell apoptosis induced by hypoxia/reoxygenation injury in vitro

Objective To explore the mechanism of apoptosis in anoxia-reoxygenation injury （H/R） on endothelial cells（ECs） and the late protective effect of catopfil. Methods To establish anoxia-reoxygenation model on human umbilical veins endothelial cell line ECV304. These ECs were randomly divided into 7 groups： different hypoxia time, different reoxygenation time, late preconditioning with catopril, catopril＋bradykinin 132 receptor inhibitor, catopril＋PKC inhibitor, eatopfil＋NOS inhibitor, catopril＋NF-KB inhibitor. Then EC apoptosis proportion and Annexin V-FITC apoptosis were observed with flow cytometry. Results The EC apoptosis proportion was increased after treatment with anoxia and anoxia-reoxygenation. Meanwhile, the changes became more obviously with the time extending. The late preconditioning with eatopril, the cell apoptosis kept normal. The anti-apoptosis effect became strengthen along with the eatopril concentration increasing. However, culturing with above four inhibitors, the anti-apoptosis protective effect was partly diminished. Conclusion Anoxia and anoxia-reperfusion induce cell apoptosis in a time-dependent manner. The late preconditioning with catopril can weaken apoptosis that is induced by anoxia-reperfusion injury in a concentration-dependent manner. Bradykinin 132 receptor, PKC activating, nitric oxide and nucleus factor are all involved in the protective effect.