胆固醇流出对ox-LDL诱导的巨噬细胞凋亡的影响

发布时间：2018-04-20 21:23

本文选题：胆固醇流出 + 氧化型低密度脂蛋白　；参考：《南华大学》2005年硕士论文

【摘要】：目的:研究胆固醇流出对ox-LDL诱导的巨噬细胞凋亡的影响。方法:用50mg/Lox-LDL与RAW264.7细胞共同孵育0,12,24,48,96小时。用50mg/Lox-LDL处理RAW264.7细胞48小时后加入不同浓度的HDL_3(50,100,200mg/L)和胆固醇流出抑制剂Brefeldin(BFA)或胆固醇流出促进剂β-cyclodextrin(β-CD)孵育24小时。油红O染色观察细胞内脂滴的形成情况;高效液相分析法检测细胞内胆固醇含量;液体闪烁计数器检测细胞内胆固醇流出;Hoechst33258观察细胞凋亡形态;流式细胞仪检测细胞凋亡。结果:用50mg/Lox-LDL与RAW264.7细胞共同孵育0,12,24,48,96小时,结果显示:RAW264.7细胞的凋亡率随着处理时间的延长而显著提高。在50mg/Lox-LDL与RAW264.7细胞共同孵育48小时出现明显的凋亡形态学改变(胞质体积缩小,胞核凝集、边缘化),油红O染色显示,细胞内的大量脂滴形成,细胞胆固醇酯含量显著增加,细胞呈典型泡沫化形态。用50mg/Lox-LDL处理RAW264.7细胞48小时后,加入不同浓度的HDL_3(50,100,200mg/L)和胆固醇流出抑制剂BFA(4μmol)或β-CD(10mmol/L)孵育24小时。结果显示:HDL_3呈浓度依赖性的促进细胞内胆固醇流出和降低细胞内的胆固醇含量,油红O染色显示,细胞内的脂滴明显减少。细胞的凋亡率也随着HDL_3浓度的升高而逐渐降
[Abstract]:Aim: to study the effect of cholesterol efflux on apoptosis of macrophages induced by ox-LDL. Methods: 50mg/Lox-LDL and RAW264.7 cells were incubated for 96 hours. RAW264.7 cells were treated with 50mg/Lox-LDL for 48 hours and incubated with different concentrations of HDL350100mg / L and cholesterol efflux inhibitor Brefeldin-BFA or cholesterol efflux enhancer 尾 -cyclodextrin (尾 -CD1) for 24 hours. Oil red O staining was used to observe the formation of lipid droplets in cells; high performance liquid chromatography (HPLC) was used to detect the content of cholesterol in cells; liquid scintillation counter was used to detect cholesterol efflux in cells; Hoechst33258 was used to observe the morphology of apoptosis; flow cytometry was used to detect apoptosis. Results: #number0# RAW264.7 cells were incubated with 50mg/Lox-LDL for 48 hours for 96 hours. The results showed that the apoptosis rate of 10% RAW264.7 cells increased significantly with the prolongation of treatment time. After 50mg/Lox-LDL was incubated with RAW264.7 cells for 48 hours, there were obvious morphological changes of apoptosis (reduced cytoplasmic volume, nuclear agglutination, marginalization, oil red O staining showed that a large number of lipid droplets were formed in the cells, and the content of cholesterol ester in the cells increased significantly. The cells showed typical foaming morphology. RAW264.7 cells were treated with 50mg/Lox-LDL for 48 hours and incubated with HCL 350100200mg / L and cholesterol efflux inhibitor BFA(4 渭 mol / L or 尾 -CDN 10mmol / L for 24 hours. The results showed that WHDL3 promoted cholesterol efflux and decreased cholesterol content in cells in a concentration-dependent manner. Oil red O staining showed that lipid droplets in the cells decreased significantly. The apoptosis rate decreased with the increase of HDL_3 concentration.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R363

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