程序性细胞死亡因子4是替代活化巨噬细胞的新分子标志物  

Programmed cell death 4 is a novel molecular marker of alternatively activated macrophages

作　　者：田稼[1] 蒋晓刚[2] 李海燕[3] 钟波[4] 张富军[2] 宁启兰[2] 韩燕[2] 杨旭东[2] 

机构地区：[1]陕西省微生物研究所,陕西省西安市710043 [2]西安交通大学医学部基础医学院生物化学与分子生物学系,陕西省西安市710061 [3]西安交通大学医学部基础医学院药理学系,陕西省西安市710061 [4]西安交通大学第二附属医院儿科,陕西省西安市710004

出　　处：《中国组织工程研究》2015年第51期8281-8288,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81370123)~~

摘　　要：背景:程序性细胞死亡因子4的表达水平与经典活化的巨噬细胞的表型负相关,但是程序性细胞死亡因子4的表达水平与巨噬细胞替代活化的关系仍不明确。目的:观察替代活化的巨噬细胞中程序性细胞死亡因子4的表达水平改变,及其对替代活化的影响。方法:用白细胞介素4、地塞米松单独刺激或联合刺激诱导NR 8383细胞的替代活化;采用qP CR检测巨噬细胞替代活化的分子标志物的表达水平,确定诱导巨噬细胞替代活化的最适条件;采用qP CR和Western blot检测替代活化的巨噬细胞模型中程序性细胞死亡因子4表达的改变;分别用大鼠程序性细胞死亡因子4高表达质粒和程序性细胞死亡因子4干扰质粒转染NR 8383细胞株,采用荧光倒置显微镜观察转染的效率,并检测程序性细胞死亡因子4的表达水平;分别检测程序性细胞死亡因子4高表达和程序性细胞死亡因子4敲低后NR 8383细胞株中巨噬细胞经典活化和替代活化的分子标志。结果与结论:(1)白细胞介素4和地塞米松联合刺激比白细胞介素4或地塞米松单独刺激可更有效的诱导巨噬细胞的替代活化;10μg/L白细胞介素4+50 nmol/L地塞米松刺激24 h可有效诱导巨噬细胞的替代活化。(2)在替代活化的巨噬细胞中程序性细胞死亡因子4的表达水平显著升高,是替代活化的分子标志。(3)程序性细胞死亡因子4高表达上调替代活化的分子标志的表达(P<0.05);而敲低程序性细胞死亡因子4可下调CD206的表达(P<0.05),并显著上调经典活化的分子标志诱导型一氧化氮合酶的表达(P<0.05)。结果表明程序性细胞死亡因子4上调是替代活化巨噬细胞的一个重要分子标志物。BACKGROUND： The expression of programmed cell death 4（PDCD4） is negatively correlated with the phenotype of classically activated macrophages, but the association between PDCD4 and alternative activation of macrophages is undefined. OBJECTIVE： To explore the changes of PDCD4 expression in alternatively activated macrophages and the effects of PDCD4 on alternative activation. METHODS： Rat macrophage cell line NR8383 was treated with interleukin-4 and dexamethasone alone or theircombination to induce alternative activation. Molecular markers of alternatively activated macrophages were detected by q PCR to identify the appropriate conditions for inducing alternative activation. The changes of PDCD4 expression in alternatively activated macrophages were detected by q PCR and western blot. NR8383 was transfected with p EGFPPDCD4 and sh PDCD4 respectively, the efficiency of transfection was evaluated by fluorescence inverted microscope, and the levels of PDCD4 were assayed by q PCR. The molecular markers of classical activation and alternative activation were detected in NR8383 with PDCD4 overexpression or knockdown. RESULTS AND CONCLUSION：（1） Combination of interleukin-4 and dexamethasone induced the alternative activation of NR8383 more efficiently than interleukin-4 or dexamethasone alone, and alternatively activated macrophages were effectively induced by 10 μg/L interleukin-4＋50 nmol/L dexamethasone for 24 hours.（2） The expression of PDCD4 increased significantly in alternatively activated macrophages, which is a novel marker of alternatively activated macrophages.（3） The overexpression of PDCD4 upregulated the expression of Arg-1 and CD206 significantly（P 〈 0.05）; the knockdown of PDCD4 down-regulated the expression of CD206（P 〈 0.05）, but up-regulated the expression of inducible nitric oxide synthase（P 〈 0.05）. In conclusion, PDCD4 upregulation is an important molecular marker of alternatively activated macrophages.

关 键 词：巨噬细胞 白细胞介素4 地塞米松 组织构建 组织工程 程序死亡因子4 替代活化 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]