余甘子总酚对RAW264.7巨噬细胞源泡沫细胞胆固醇代谢的影响研究

Effects of Total Phenols of PHYLLANTHI FRUCTUS on Cholesterol Metabolism of RAW264.7 Macrophage-Derived Foam Cells

目的:观察余甘子总酚对RAW264.7巨噬细胞源泡沫细胞胆固醇代谢的影响。方法:80μg/mL氧化修饰型低密度脂蛋白(ox-LDL)诱导RAW264.7巨噬细胞泡沫化,通过油红O染色、胆固醇含量鉴定泡沫细胞模型,用不同浓度的余甘子总酚处理RAW264.7泡沫细胞后,测定细胞内脂质及胆固醇含量;采用q-PCR和Western blotting方法检测胆固醇代谢相关基因和蛋白表达。结果:与正常对照组比较,模型对照组细胞内脂滴增多,总胆固醇(TC)、胆固醇酯(CE)含量显著增多(P<0.01),清道夫受体(Cd36、Sra1)基因表达明显上调(P<0.05或P<0.01),CD36、SR-A1、ABCA1蛋白表达明显上调(P<0.05或P<0.01);与模型对照组比较,余甘子总酚75、150、300μg/mL组可剂量依赖性降低细胞内TC、CE含量(P<0.01),明显下调Cd36、Sra1基因及蛋白表达(P<0.05或P<0.01),余甘子总酚150、300μg/mL组ATP结合盒转运体A1(Abca1)基因表达显著上调(P<0.01),余甘子总酚300μg/mL组ATP结合盒转运体G1(Abcg1)基因及ABCA1蛋白表达明显上调(P<0.05)。结论:余甘子总酚可通过调节CD36、SRA1、ABCA1介导的胆固醇转运而影响RAW264.7巨噬细胞源泡沫细胞的胆固醇代谢,提示余甘子总酚可通过改善巨噬细胞胆固醇代谢而起到抗动脉粥样硬化的作用。

Objective:To observe the effects of total phenols of PHYLLANTHI FRUCTUS on cholesterol metabolism of RAW264.7 macrophage-derived foam cells.Methods:RAW264.7 macrophages were foamed by 80μg/mL oxidized low-density lipoprotein(ox-LDL).The foam cell model was identified by oil red O staining and cholesterol level.After RAW264.7 foam cells were treated with total phenols of PHYLLANTHI FRUCTUS of different concentrations,the lipid and cholesterol content in cells was determined.The expression levels of genes and proteins related to cholesterol metabolism were detected by q-PCR and Western blot.Results:Compared with the normal group,the model group showed increased lipid droplets,elevated content of total cholesterol(TC)and cholesterol ester(CE)(P<0.01),and up regulated gene expression levels of scavenger receptors(Cd36 and Sra1)(P<0.05 or P<0.01),and protein expression levels of CD36,SR-A1,and ABCA1(P<0.01 or P<0.05).Compared with the conditions in the model group,75,150,and 300μg/mL total phenols of PHYLLANTHI FRUCTUS decreased intracellular TC and CE levels in a dose-dependent manner(P<0.01)and down-regulated the gene and protein expression of Cd36 and Sra1(P<0.05 or P<0.01).Additionally,150 and 300μg/mL total phenols of PHYLLANTHI FRUCTUS up regulated the gene expression level of ATP-binding cassette transporter A1(Abca1)(P<0.01),and 300μg/mL total phenols of PHYLLANTHI FRUCTUS up regulated the gene expression of ATP-binding cassette transporter G1(Abcg1)and protein expression of ABCA1(P<0.05).Conclusion:Total phenols of PHYLLANTHI FRUCTUS can affect cholesterol metabolism of RAW264.7 macrophage-derived foam cells by regulating cholesterol transport mediated by CD36,SRA1,and ABCA1,suggesting that total phenols of PHYLLANTHI FRUCTUS can play an anti-atherosclerotic effect by improving cholesterol metabolism of macrophages.