大鼠脑微血管内皮细胞体外缺血再灌注模型的建立及他汀的保护作用

目的 建立大鼠脑微血管内皮细胞体外缺血再灌注模型,并观察辛伐他汀、洛伐他汀的保护作用.方法 原代分离培养SD大鼠脑微血管内皮细胞,以无糖Krebs溶液再复氧复糖模拟体外缺血再灌注损伤,并实施辛伐他汀、洛伐他汀预处理干预.观察各组细胞形态,测定各组细胞上清中内皮型一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)活性,利用CCK-8检测各组细胞增殖活性.结果 氧糖剥夺(Krebs液) 缺氧12h再复氧复糖4h模型可成功模拟大鼠脑微血管内皮细胞缺血再灌注损伤.辛伐他汀、洛伐他汀预处理后,脑微血管内皮细胞eNOS活性增强、iNOS活性降低,细胞增殖活性提高.结论 辛伐他汀、洛伐他汀具有显著的脑缺血再灌注损伤保护作用.

Establishment of an in vitro injury model of ischemia-reperfusion with rat brain microvascular endothelial cell and the protective effect of statins

Objective To establish an in vitro injury model of ischemia-reperfusion with rat brain microvascular endothelial cells(BMECs) and observe the protective effect of statins.MethodsSD rat BMECs were isolated and cultured by enzyme digestion method,BMECs were subjected to the oxygen-glucose deprivation(OGD)(Krebs solution) and recovery of oxygen-glucose,which simulated in vitro ischemia-reperfusion injury,and pretreated with simvastatin and lovastatin.The activity of nitric oxide synthase(NOS) was determined in cell medium.The BMECs' multiplication activity was detected by Cell Counting Kit-8.ResultsBMECs were impaired after OGD for 12 hours and recovery of oxygen-glucose for 4 hours.The BMECs' multiplication activity with pretreatment of statins was significantly higher than control group(P <0.01).ConclusionSimvastatin and lovastatin can obviously protect BMECs from an in vitro ischemia-reperfusion injury.