摘要
目的观察脂肪因子chemerin是否调节巨噬泡沫细胞形成,以及对巨噬泡沫细胞CD36、ATP结合盒转运子A1(ATP-binding cassette transporter A1,ABCA1)表达和炎症因子释放的影响。方法 100nmol/L佛波酯(phorbolmyristate acetate,PMA)诱导THP-1单核细胞分化为巨噬细胞,随机分组,Chemerin不同浓度和作用时间下联合相同浓度(50μg/mL)氧化型低密度脂蛋白与THP-1源性巨噬细胞共孵育。油红O染色观察细胞内脂滴变化,Western blot和实时PCR检测CD36、ABCA1的mRNA和蛋白水平的表达。ELISA法检测培养液中炎症因子———肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)和白细胞介素10(interleukin-10,IL-10)的浓度,酶法检测细胞内胆固醇含量。结果Chemerin诱导巨噬细胞内脂滴蓄积,下调CD36和ABCA1mRNA及蛋白的表达,增加培养液中促炎因子TNF-α的释放而降低抗炎因子IL-10的释放,提高细胞内胆固醇的含量,且有浓度和时间依赖性。结论 Chemerin通过下调CD36和ABCA1基因的mRNA及蛋白的表达,诱导THP-1巨噬细胞内脂滴蓄积和泡沫细胞形成;ABCA1表达下调可能与促炎因子TNF-α释放增加和抗炎因子IL-10释放减少有关。
Objective To determined if chemerin can modulate macrophage foam cells formation,to investigate its effect on the expression of CD36 and ATP-binding cassette transporter A1（ABCA1）,and the release of inflammation cytokines in macrophage foam cells.Methods THP-1 cells were induced into macrophages by 100 nmol/L phorbol myristate acetate（PMA）for 48 h.The THP-1-derived macrophages were co-cultured with chemerin with different concentrations and durations in combination with 50 μg/mL oxidized low-density lipoprotein.Intracellular lipid droplets were observed by oil red O staining.The expression levels of ABCA1 and CD36 mRNA and protein were detected by using Western blot and real time-polymerase chain reaction（RT-PCR）.Inflammatory cytokines [tumor necrosis factor-alpha（TNF-α）and interleukin-10（IL-10）] concentrations in medium were determined by using ELISA.The contents of cholesterol were determined by using enzyme fluorescence.Results Chemerin induced lipid droplet accumulation,increased intracellular cholesterol contents,reduced the expression of CD36 and ABCA1 mRNA and protein in THP-1-derived macrophages,increased the release of pro-inflammatory cytokines TNF-α,and decreased the release of anti-inflammatory cytokines IL-10 in a dose-and time-dependent manner.Conclusion Chemerin increased THP-1-derived macrophages lipid droplet accumulation and foam cells formation by decreasing the expression of CD36 and ABCA1 mRNA and protein.The down-regulation of ABCA1 expression may be associated with the increased release of pro-inflammatory cytokines TNF-α and the decreased release of anti-inflammatory cytokines IL-10.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期27-31,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30872541)
博士点基金新教师项目资助课题(No.200804871116)
