SCARF1在人THP-1单核源性巨噬细胞抗烟曲霉菌刺激中的作用  

The Role of SCARF1 in THP1 Derived Macrophage Defense against Aspergillus Fumigatus

作　　者：鄢雯[1] 李囡[1] 刘立新[1] 夏山[1] 刘扬[1] 

机构地区：[1]首都医科大学燕京医学院,北京101300

出　　处：《现代生物医学进展》2016年第25期4815-4818,共4页Progress in Modern Biomedicine

基　　金：北京高等学校青年英才项目(YETP1679);首都医科大学自然科学基金项目(2014QD03)

摘　　要：目的:运用干扰腺病毒沉默THP1细胞中SCARF1基因研究其在体外抗烟曲霉中的作用。方法:用灭活的烟曲霉分生孢子(1×105 CFU/m L)于不同时间点处理THP-1细胞,RT-PCR分别检测SCARF1和TNF-αm RNA的表达;将Ad-si RNA-SCARF1转导细胞24 h后给予烟曲霉孢子刺激24 h,通过RT-PCR法检测细胞中TNF-αm RNA表达,Western blot法检测细胞中SCARF1表达以及NF-κB通路相关信号分子的活性。结果:RT-PCR证实烟曲霉孢子刺激能时间依赖性增强THP1细胞中SCARF1和TNF-α表达;Western法证实与Ad-GFP组比较Ad-si RNA-SCARF1组SCARF1的表达量显著降低(P<0.05),沉默效率为71%;与Ad-GFP组比较,Ad-GFP+Af组NF-κB亚单位p65的磷酸化水平明显升高(P<0.05),在Ad-si RNA-SCARF1+Af组,磷酸化p65的产生明显减少,SCARF1沉默后细胞因子TNF-α的分泌明显减少。结论:烟曲霉孢子刺激能诱导巨噬细胞SCARF1的表达增加,诱导信号分子NF-κB的活化,导致相应的细胞因子分泌增加,从而在巨噬细胞抗烟曲霉中发挥作用。Objective： To study the role of SCARF1 against Aspergillus fumigatus in THP1 cells through knockdown of SCARF1 by adenoviruses expressing sh RNA against SCARF1. Methods： The expression of SCARF1 and TNF-α gene of THP1 cells were measured by RT-PCR after treatment with 1×105CFU / m L inactivated Aspergillus fumigatus spores for different time lengths. After knockdown of SCARF1 expression with adenoviruses expressing sh RNA against SCARF1, macrophages were stimulated for 24 h by Aspergillus fumigatus conidia. The expression levels of TNF-α m RNA, SCARF1 protein and signal molecules of NF-κB pathway were examined by RT-PCR and Western blot. Results： Aspergillus fumigatus spores stimulation can increase the expression of SCARF1 and TNF-α in THP1 cells time-dependently. Silencing efficiency of Ad-si RNA-SCARF1 was up to 71 % in macrophages（P〈0.05）. Compared with the Ad-GFP group,-p65 phosphorylation was increased significantly in Ad-GFP＋Af group（P〈0.05）; Compared with Ad-GFP＋Af group, the p65 phosphorylation and secretion of cytokine TNF-α were lowered significantly in SCARF1（RNAi）＋Af group.Conclusion： SCARF1 in macrophages was activated by Aspergillus fumigatus conidia and exerted the effect of anti-Aspergillus fumigatus spores stimulation through stimulating NF-κB signaling pathway and releasing pro-inflammatory cytokines.

关 键 词：SCARF1 烟曲霉菌 THP-1细胞 NF-ΚB 

分 类 号：R379[医药卫生—病原生物学] R-33[医药卫生—基础医学]