氧化芍药苷对泡沫细胞胆固醇流出的影响

Effect of Oxypaeoniflora on cholesterol efflux from foam cells

摘要:

分析氧化芍药苷对THP-1 巨噬细胞源性泡沫细胞胆固醇流出的影响。方法THP-1 细胞加入160 nmol/L 佛波酯（PMA）24 h，再加入50 μg/ml 乙酰化低密度脂蛋白（ac-LDL）建立泡沫细胞模型，CCK-8 法测定氧化芍药苷的细胞毒性，同位素标记法测定各处理组泡沫细胞胆固醇流出率。共聚焦显微镜观察泡沫细胞内脂肪分化相关蛋白（ADFP）表达变化，Western blot分析泡沫细胞中ADFP、三磷酸腺苷结合盒转运体A1（ABCA1）和过氧化物酶体增殖物激活受体α（PPARα）蛋白表达变化。结果成功建立THP-1 巨噬细胞源性泡沫细胞模型。氧化芍药苷毒性分析表明，质量浓度在200.0μg/ml 以下无细胞毒性。150.0μg/ml 与200.0μg/ml氧化芍药苷处理泡沫细胞，胆固醇流出率分别为（10.317±0.508）%与（11.265±0.713）%，与apoA-1组相比，差异有统计学意义（p <0.05），均高于apoA-1 组。氧化芍药苷处理的泡沫细胞内ADFP 荧光明显减弱。Western blot分析泡沫细胞ADFP 表达量降低28%。氧化芍药苷处理组PPARα 表达量增加51%，ABCA1 表达量增加45%。结论实验表明氧化芍药苷可通过上调PPARα增加ABCA1 表达，进而促进泡沫细胞胆固醇流出。

Abstract:

To analyze the effect of Oxypaeoniflora on cholesterol efflux from THP-1 macrophagederived foam cells. Methods THP-1 cells were treated with 160 nmol/L phorbol-1-myristate-13-acetate (PMA) for 24 h, then 50 μg/ml acetylated low-density lipoprotein (ac-LDL) to establish foam cell model. Cytotoxicity of Oxypaeoniflora was assessed by CCK-8. Cholesterol efflux rate of the foam cells in each treatment group was evaluated by isotope labeling method. The expression of adipose differentiation related protein (ADFP) in the foam cells was observed by confocal laser scanning microscopy. The expression changes of ADFP, ATP binding cassette transporter A1 (ABCA1) and peroxisome proliferator-activated receptor alpha (PPARα) in the foam cells were detected using Western blot. Results The THP-1 macrophage-derived foam cell model was successfully established. At the concentration of lower than 200.0 μg/ml, Oxypaeoniflora was not cytotoxic. When the Oxypaeoniflora concentrations were 150.0 μg/ml and 200.0 μg/ml, the cholesterol efflux rates were about (10.317 ±0.508)% and(11.265 ±0.713)% respectively, which were increased compared with the control group (p < 0.05). Under microscope, the fluorescence of ADFP in the Oxypaeoniflora groups was obviously decreased. Western blot indicated that the expression of ADFP was decreased by 28%, while the expressions of PPARαand ABCA1 were increased by 51% and 45% respectively. Conclusions The results state clearly Oxypaeoniflora can increase the expression of ABCA1 by up-regulation of PPARα expression and then promote the cholesterol efflux from foam cells.