Nifedipine对ApoE--小鼠RAW264.7巨噬细胞源性胆固醇逆向转运影响的研究.docVIP

Nifedipine对ApoE-/-小鼠RAW264.7巨噬细胞源性胆固醇逆向转运影响的研究 王 婵，张 倩，沙马阿直，宋治远 (400038 重庆，第三军医大学西南医院心血管内科，重庆市介入心脏病学研究所) [] 目的 探讨硝苯地平对ApoE-/-小鼠RAW264.7巨噬细胞源性胆固醇逆向转运的影响。方法 8周龄左右ApoE-/-小鼠16只，用高脂饲料喂养作为模型，按完全随机分组法分为对照组（n=8）和硝苯地平组（n=8）。将[3H]-胆固醇标记的荷脂RAW264.7小鼠巨噬细胞注入小鼠腹腔，24、48h时取血液标本，酶法检测血浆总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的浓度。处死后取小鼠肝脏和小肠，实时定量PCR检测肝脏及小肠ABCA1、ABCG1、SR-BI、LXRα基因的mRNA表达，Western blot检测其蛋白表达。同时，收集48h小鼠粪便，进行闪烁液计数，分别比较两组[3H]的含量。结果 与对照组相比，经硝苯地平处理组TC降低0.8%（P0.05），其余血脂水平无明显变化；血浆24、48h cpm值较对照组分别增加37.02%、22.72%（P0.05），粪便[3H]cpm值较对照组增加43.18%（P0.05）；硝苯地平组ABCA1、ABCG1、SR-BI、LXRα基因的mRNA和蛋白的表达基因表达均有明显升高。结论 硝苯地平通过增加LXRα及其下游基因ABCA1、ABCG1、SR-BI的表达，从而增强ApoE-/-小鼠巨噬细胞源性胆固醇逆向转运的效果。 [关键词] 胆固醇逆向转运；巨噬细胞；硝苯地平；ApoE-/-小鼠 [中图法分类号] [文献标志码] A The effect of Nifedipine on Macrophage Reverse Cholesterol Transport in ApoE-/- Mice Wang Chan, Zhang Qian, ShaMaAzhi, Song Zhiyuan(Department of Cardiovascular Diseases，Southwest Hospital，Third Military Medical University, Chongqing 400038，China) [Abstract] Objective To investigate whether nifedipine increases macrophage reverse cholesterol transport in ApoE-/- mice and the expression of related genes. Methods Sixteen eight-week male ApoE-/- mice(fed with high fat and cholesterol) were randomly divided into two groups: control group and nifedipine group.RAW264.7 macrophages were loaded with cholesterol by incubation with acetylated-LDL, labeled with[3H]-cholesterol, and then intraperitoneally injected into mice. Blood was collected in 24h、48h after injection, tissues and feces were collected in 48hours, and all concentrations of plasma lipids(TC, TG, LDL and HDL)were measured by Biochemistry Analyzer. Blood and feces were analyzed for tracer counts using a liquid scintillation counter. Related mRNA and protein levels of several well-characterized proteins involved in reverse cholesterol transportation, including ABCA1, ABCG1, SR-BI and LXRα, using quantitative real-time PCR, Western blot tech