亲环素A对THP-1源性泡沫细胞ABCA1表达及胆固醇流出的影响及机制  

Effects of cyclophilin A on ABCA1 expression and cholesterol efflux in THP-1 derived macrophages and its mechanism

作　　者：唐小二 张敏[2] 李靓[2] 谢巍[2] 龚朵 夏晓丹[2] 成海鹏 赵真旺 陈凌燕 唐朝克[2] TANG Xiao-Er ZHANG Min LI Liang XIE Wei GONG Duo XIA Xiao-Dan CHENG Hai-Peng ZHAO Zhen- Wang CHEN Ling-Yan TANG Chao-Ke(Department of Pathophysiology, Medical College, Shaoyang University, Shaoyang, Hunan 422000, China Institnte of Cardiovascular Diseases, University of South China ＆ Key Laboratory for Atherosclerology of Hunan Province ＆ Medical Re- search Center, University of South China ＆ Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study, Hengyang, Hunan 421001, China)

机构地区：[1]邵阳学院医学院病理生理学教研室,湖南省邵阳市422000 [2]南华大学心血管疾病研究所动脉硬化学湖南省重点实验室南华大学医学研究中心分子靶标湖南省协同创新中心,湖南省衡阳市421001

出　　处：《中国动脉硬化杂志》2017年第9期879-884,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81570408;81370377和81470569);南华大学研究生创新课题(2016XCX04)

摘　　要：目的观察亲环素A(CypA)对THP-1源性泡沫细胞三磷酸腺苷结合盒转运体A1(ABCA1)表达和胆固醇流出的影响及机制。方法 160 nmol/L佛波酯诱导人THP-1单核细胞分化为巨噬细胞,与50 mg/L氧化型低密度脂蛋白(ox-LDL)共孵育,使其荷脂形成泡沫细胞,常规体外培养细胞;实验分为对照组和Cyp A处理组;液体闪烁计数仪检测细胞胆固醇流出水平,高效液相色谱检测细胞内脂质成分,实时荧光定量PCR和Western blot检测ABCA1表达,Western blot检测核因子κB(NF-κB)核转位水平;NF-κB抑制剂小白菊内酯抑制NF-κB活化;脂质体2000转染CD147 siRNA至THP-1源性泡沫细胞。结果 CypA显著抑制THP-1源性泡沫细胞胆固醇流出,促进NF-κB核转位,下调ABCA1表达;NF-κB抑制剂小白菊内酯拮抗CypA对ABCA1表达及胆固醇流出的抑制作用;用siRNA干扰CD147后,显著抑制Cyp A诱导的NF-κB核转位,上调ABCA1表达,促进胆固醇流出。结论 CypA可能通过CD147激活NF-κB,下调ABCA1表达,抑制THP-1源性泡沫细胞胆固醇流出。Aim To investigate the effects and mechanisms of cyclophilin A （CypA） on the expression of ATP binding cassette trasporter A1 （ABCA1） and cholesterol efflux in THP-1 derived macrophages. Methods THP-1 cells were incubated with 160 nmol/L phorbol ester to transform into macrophages. THP-1 derived macrophages were exposed to 50 mg/L oxidized low density lipoprotein （ox-LDL） and cultured with typical approaches. Liquid scintillation counter was used to determine the efficiency of cholesterol efflux. Lipids contents were tested with high performance liquid chro- matogram （HPLC）. Real-time PCR and Western blot were used to quantify the expression of ABCA1 and nuclear factor- KB （NF-KB） nuclear translocation. Parthenolide was used as the specific inhibitor of NF-KB. CD147 was silenced with siRNA. Results CypA significantly reduced cholesterol efflux, promoted NF-KB nuclear translocation and downregu- lated ABCA1 expression. NF-KB inhibitor parthenolide interfered CypA-prevented ABCA1 expression and cholesterol ef- flux. After CD147 were silenced with siRNA, CypA inhibited NF-KB nuclear translocation, upregulated ABCA1 expression and promoted cholesterol efflux. Conclusion CypA prevented THP-1 derived macrophages cholesterol efflux by activing NF-KB in a CD147 dependent manner and downregulating ABCA1 expression.

关 键 词：亲环素A 三磷酸腺苷结合盒转运体A1 胆固醇流出 核因子κB CD147 

分 类 号：R363[医药卫生—病理学]