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目的研究阿托伐他汀对脂多糖(Lipopolysaccharide,LPS)诱导的小鼠巨噬细胞RAW264.7脂代谢相关蛋白表达及胆固醇流出的影响,并探讨其可能的作用机制。方法用不同浓度LPS刺激RAW264.7细胞,在不同时间点以Western blot方法检测脂质代谢相关蛋白ABCA1、ABCG1、SR-B1、PCSK9和LDLR的表达,从而确定LPS最佳作用浓度和时间。Western blot检测不同浓度阿托伐他汀对50ng/m L LPS诱导的上述脂代谢相关蛋白表达变化的影响,激光共聚焦观察BODIPY荧光标记的胆固醇流出状况。结果在RAW264.7细胞中,LPS呈浓度和时间依赖性抑制ABCA1、ABCG1、SR-B1和LDLR蛋白的表达,对PCSK9蛋白表达无显著影响。阿托伐他汀浓度依赖性的上调LPS诱导的ABCA1、ABCG1、SR-B1和LDLR蛋白表达降低,上调PCSK9蛋白的表达,从而增加胆固醇流出,减少细胞内胆固醇聚集。结论阿托伐他汀干预能够改善LPS引起的脂代谢相关蛋白表达并增强胆固醇流出,该作用可能是其抗动脉粥样硬化机制之一。
Objective To investigate the effect of atorvastatin on lipopolysaccharide (LPS) -induced RAW264.7 lipid metabolism-related protein expression and cholesterol efflux in murine macrophages and to explore its possible mechanism. Methods RAW264.7 cells were stimulated with different concentrations of LPS, and the expression of lipid metabolism-related proteins ABCA1, ABCG1, SR-B1, PCSK9 and LDLR were detected by Western blot at different time points to determine the optimal concentration and time of LPS. Western blot was used to detect the effects of different concentrations of atorvastatin on the expression of lipid metabolism-related proteins induced by LPS at a dose of 50ng / m L, and the efflux of BODIPY-labeled cholesterol was observed by confocal laser scanning microscope. Results LPS inhibited the expression of ABCA1, ABCG1, SR-B1 and LDLR protein in a concentration-and time-dependent manner in RAW264.7 cells without any significant effect on the expression of PCSK9 protein. Atorvastatin up-regulates LPS-induced decrease of ABCA1, ABCG1, SR-B1 and LDLR protein expression and up-regulates the expression of PCSK9 protein, thereby increasing cholesterol efflux and decreasing intracellular cholesterol accumulation. Conclusions Atorvastatin treatment can improve LPS-induced lipid metabolism-related protein expression and enhance cholesterol efflux, which may be one of the mechanisms of anti-atherosclerosis.