辛伐他汀对不同氧供条件下大鼠脑微血管内皮细胞跨内皮电阻及MMP-9表达的影响

Effect of Simvastatin on Endothelial Permeability and Expression of Matrix metallproteinase-9 in Rat Brain Microvascular Endothelial Cell under Different Oxygen Supply Conditions

目的:观察在常氧、短暂缺氧、持续缺氧及复氧条件下,辛伐他汀对大鼠脑微血管内皮细胞(BMECs)跨内皮细胞电阻(TEER)及基质金属蛋白酶-9(MMP-9)表达的影响。方法:原代分离培养SD大鼠BMECs,随机分为常氧组、短暂缺氧组、持续缺氧组、复氧组,各组均实施辛伐他汀0.1μmol/L、1.0μmol/L、10μmol/L3种浓度预处理。观察各组细胞形态学及增殖活性变化;采用TEER评估各组通透性;免疫细胞荧光化学法测定MMP-9蛋白表达。结果:短暂缺氧、持续缺氧及复氧条件下大鼠BMECs细胞受损逐渐加重,MMP-9表达逐渐增加,TEER及细胞增殖活性逐渐下降。辛伐他汀干预能不同程度提高TEER并抑制MMP-9蛋白的表达,使细胞增殖活性提高。但这种抑制MMP-9的作用对短暂缺氧大鼠BMECs的TEER值及细胞增殖活性无显著改善。结论:辛伐他汀能通过抑制MMP-9蛋白的表达来提高持续缺氧及复氧条件下大鼠BMECs的TEER,而对短暂缺氧后TEER的改善作用不明显。

Objective:To observe the influence of Simvastatin on transendothelial electrical resistance(TEER) and expression of matrix metallproteinase-9(MMP-9) in rat brain microvascular endothelial cells(BMECs) under normoxia,transient hypoxia,continuous hypoxia or reoxygenation conditions.Method:SD rat BMECs were isolated and cultured by enzyme digestion method,and randomly assigned into 4 groups:normoxia,transient hypoxia,continuous hypoxia and reoxygenation.Different concentrations(0.1,1.0,10 mol/L) of Simvastatin were added to each group.BMECs hypoxia/reoxygenation model was built in vitro.The cell proliferation activity was detected with CCK-8 kit.Transendothelial electrical resistance(TEER) techniques were employed to measure changes in permeability across the BMEC monolayer.The expression of MMP-9 in BMECs was detected by cell immunofluorescence technique.Results:Hypoxia/reoxygenation markedly increased the expression of MMP-9 and permeability in BMECs.Simvastatin reduced the expression of MMP-9 and increased the TEER of the BMEC monolayer in different degree for each group.Such effect in transient hypoxia,continuous hypoxia and reoxygenation group is obvious.Conclusion:The inhibition effect of Simvastatin on MMP-9 expression contributes to the reduction of permeability of BMECs under hypoxia or reoxygenation conditions.