摘要
目的:探讨瑞舒伐他汀改善缺氧/复氧引起的小鼠脑微血管内皮细胞(brain microvascular endothelial cells,BMECs)损伤的作用及其可能机制。方法:原代分离培养BALB/c小鼠BMECs,利用氧糖剥夺/复氧模拟体外缺血再灌注损伤,以瑞舒伐他汀处理干预。观察细胞形态,通过MTT实验测定BMECs细胞的活力,荧光染料羧基荧光素二乙酸酯琥珀酰亚胺脂(carboxyfluorescein diacetate succinimidyl ester,CFDA-SE)细胞增殖检测试剂盒观察造模及用药后细胞数目百分比的变化;免疫荧光染色检测细胞凋亡相关蛋白cleaved caspase-3的水平; Western blot法检测Bcl-2、Bax、基质金属蛋白酶(matrix metalloproteinase,MMP) 2、MMP9、磷酸化核因子κB(phosphorylated nuclear factor kappa B,p-NF-κB)、磷酸化P38丝裂原活化蛋白激酶(phosphorylated P38 mitogen-activated protein kinase,p-P38)和磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase,p-JNK)的蛋白水平。结果:浓度为10μmol/L的瑞舒伐他汀可抑制氧糖剥夺/复氧诱导的BMECs存活率减低现象,同时维持BMECs结构。此外,瑞舒伐他汀能够显著抑制cleaved caspase-3、MMP2、MMP9、p-NF-κB、p-P38及p-JNK的蛋白水平(P <0. 05),上调氧糖剥夺/复氧导致的Bcl-2/Bax降低(P <0. 05)。结论:瑞舒伐他汀可通过降低凋亡相关蛋白及基质金属蛋白酶的表达,对抗缺氧/复氧引起的BMECs损伤,提示其在缺血再灌注损伤时维持血脑屏障完整性方面具有潜在应用价值。
AIM:To explore the effect of rosuvastatin on the oxygen-glucose deprivation(OGD)/reoxygenation induced injury of cerebral microvascular endothelial cells(BMECs).METHODS:BMECs derived from BALB/c mice were isolated and cultured.BMECs were pretreated with rosuvastatin,followed by OGD for 3 h or 6 h and reoxygenation for 24 h.The morphological changes of BMECs were observed under light microscope.MTT assay was used to measured the cell viability,and carboxyfluorescein diacetate succinimidyl ester(CFDA-SE)staining was used to assess the proliferation of BMECs.The protein levels of cleaved caspase-3 was observed by immunofluorescence staining.The protein levels of Bcl-2,Bax,matrix metalloproteinase(MMP)2,MMP9,phosphorylated nuclear factor kappa B(p-NF-κB),phosphorylated P38 mitogen-activated protein kinase(p-P38)and phosphorylated c-Jun N-terminal kinase(p-JNK)were determined by Western blot.RESULTS:Rosuvastatin at 10μmol/L improved the viability of the BMECs with OGD/reoxygenation-induced damage,and maintained the structure of BMECs.Moreover,rosuvastatin significantly prohibited the protein levels of cleaved caspase-3,MMP2,MMP9,p-NF-κB,p-P38 and p-JNK,and up-regulated the ratio of Bcl-2/Bax(P<0.05).CONCLUSION:Rosuvastatin reduces OGD/reoxygenation-induced injury of BMECs by inhibiting the expression of apoptosis-related proteins and MMPs,suggesting that rosuvastatin has potential value for the maintenance of blood-brain barrier.
作者
王圆圆
白雅
马晨超
郭超
秦娜
魏东
WANG Yuan-yuan;BAI Ya;MA Chen-chao;GUO Chao;QIN Na;WEI Dong(Xijing Hospital,The Fourth Military Medical University,Xi an 710032,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第10期1796-1804,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81403134)
关键词
瑞舒伐他汀
血脑屏障
氧糖剥夺
脑微血管内皮细胞
细胞凋亡
Rosuvastatin
Blood-brain barrier
Oxygen-glucose deprivation
Brain microvascular endothelial cells
Apoptosis
