荷叶碱对巨噬细胞源性泡沫细胞ABCA1表达与胆固醇流出的影响及机制

Nuciferine promotes ABCA1 expression and cholesterol efflux in THP-1 macrophage-derived foam cells and its mechanism

目的观察荷叶碱（NF）对巨噬细胞源性泡沫细胞ATP结合盒转运体A1（ABCA1）表达和胆固醇流出的影响及机制。方法体外培养人源性THP-1单核细胞,使用佛波酯（160 nmol/L）对其进行诱导,促进其分化为巨噬细胞,并在50 mg/L氧化低密度脂蛋白处理下,使其荷脂形成泡沫细胞,进行常规体外培养细胞。高效液相色谱检测泡沫细胞内脂质成分,液体闪烁计数仪检测泡沫细胞胆固醇流出的水平;实时荧光定量PCR和Western blot检测ABCA1表达情况。PPARγ抑制剂GW9662和LXRα抑制剂GGPP分别处理泡沫细胞,通过荧光定量PCR和Western blot检测荷叶碱对泡沫细胞PPARγ、LXRα和ABCA1表达的影响。结果荷叶碱显著减少泡沫细胞内的脂质成分,增加ABCA1表达与胆固醇流出水平。PPARγ抑制剂GW9662处理细胞后,显著降低了PPARγ、LXRα和ABCA1的表达。泡沫细胞在LXRα抑制剂GGPP处理下,也明显降低了LXRα和ABCA1表达。结论荷叶碱通过PPARγ/LXRα途径上调巨噬细胞源性泡沫细胞ABCA1的表达,促进胆固醇流出,减少细胞内脂质蓄积。

Aim To investigate the effects and mechanisms of nuciferine（NF） on the expression of ATP binding cassette transporter A1（ABCA1） and cholesterol efflux in THP-1 macrophages-derived foam cells. Methods THP-1 cells were differentiated into macrophages by 160 nmol/L phorbol myristate acetate. THP-1 derived macrophages were incubated with 50 mg/L oxidized low density lipoprotein to induce foam cells and cultured with typical approaches. Lipids contents were tested with high performance liquid chromatogram. Liquid scintillation counter was used to determine the efficiency of cholesterol efflux. Real-time PCR and Western blot were used to quantify the expression of ABCA1. The THP-1 macrophages derived foam cells were pretreated with peroxisome proliferator-activated receptor γ（PPARγ） inhibitor GW9662 or LXRα inhibitor GGPP to detect the expression of ABCA1 by real-time quantitative PCR and Western blot.Results NF obviously reduced the intracellular Lipids contents,increased the ABCA1 expression and cholesterol efflux in THP-1 macrophages derived foam cells. PPARγ inhibitor GW9662 and LXRα inhibitor GGPP both interfered NFpromoted ABCA1 expression. Conclusion NF up-regulates the expression of ABCA1 by PPARγ/LXRα pathway to promote the cholesterol efflux and reduce the lipid accumulation in THP-1 macrophages derived foam cells.