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-  2018 

FABP5促进RAW264.7细胞氧化LDL的初步研究 Preliminary Study of FABP5 Promotes LDL's Oxidation in RAW264.7 Macrophages

Keywords: 脂肪酸结合蛋白5,磷脂酶A2,不饱和脂肪酸,脂筏,低密度脂蛋白氧化

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Abstract:

目的:探究RAW264.7细胞中脂肪酸结合蛋白5(FABP5),磷脂酶A2(PLA2),不饱和脂肪酸(UFAs)在低密度脂蛋白(LDL)氧化中的作用。方法:培养RAW264.7细胞,利用UFAs中的不同浓度的油酸(OA)、亚油酸(ALA)、花生四烯酸(AA)刺激后利用Western Blot检测细胞FABP5蛋白质水平;用LDL刺激RAW264.7细胞不同时间,分别用PLA2总活性分光光度法定量检测试剂盒测定细胞内PLA2活性变化,Real-time PCR及Western Blot测定FABP5基因的表达;LDL刺激细胞后分别测定脂质筏和非脂质筏FABP5蛋白质水平,探究FABP5的转位。结果:不同浓度的OA,ALA,AA长时间刺激RAW264.7细胞后,FABP5的蛋白表达量出现不同的变化;LDL短时间刺激小鼠巨噬细胞RAW264.7后,细胞内PLA2活性升高;LDL刺激RAW264.7细胞较长时间后,FABP5的mRNA和蛋白质水平均较对照组上升;提取脂质筏后发现,LDL刺激15min的巨噬细胞的脂筏中FABP5水平较对照组的升高,而非脂筏部分则出现相反的结果。结论:推测LDL刺激会激活PLA2,而PLA2水解甘油磷脂产生UFAs,这些UFAs招募并结合FABP5,激发氧化应激通路,促进LDL的氧化

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