摘要
目的探讨长链酰基辅酶A脱氢酶(Acyl-CoA dehydrogenase,long chain,ACADL)对巨噬细胞分泌炎症因子的影响。方法在体外诱导的小鼠骨髓来源巨噬细胞中加入脂多糖(LPS)以及白细胞介素4(IL-4)分别诱导M1和M2极化,在极化0、12、24、36、48 h时用Western blot检测ACADL在M1和M2极化中的蛋白表达量。构建ACADL-pEGFP-N1质粒,使用G418筛选和鉴定过表达ACADL的RAW264.7细胞稳定株。LPS刺激过表达ACADL稳定株和对照组细胞4 h后,通过ELISA和qRTPCR检测TNF-α、IL-6、IL-10的表达情况。在RAW264.7细胞中加入beta氧化抑制因子etomoxir预处理,ELISA检测TNF-α、IL-6、IL-10的表达情况。结果在巨噬细胞极化过程中ACADL蛋白量先减少再增加;过表达ACADL的细胞中,IL-6和IL-10分泌量及mRNA水平均显著高于对照组(P<0.001):beta氧化抑制因子etomoxir能够减少RAW264.7细胞分泌炎症因子。结论ACADL能增强巨噬细胞IL-6和IL-10的分泌,其作用机制可能是通过beta氧化影响炎性细胞因子的表达。
Macrophages are an important part of the innate immune system, which respond to different stimulating factors and differentiate into different phenotypes to participate in the normal physiological responses of the body and the occurrence and development of diseases. This study was performed to investigate the effects of mouse Acyl-CoA dehydrogenase, long chain(ACADL) on the secretion of inflammatory factors by macrophages.Lipopolysaccharide(LPS) and interleukin-4(IL-4) were added to mouse bone marrow-derived macrophages in vitro to induce M1 and M2 polarization, respectively. The protein expression of ACADL during M1 and M2 polarization was detected by Western blotting at 0, 12, 24, 36 and 48 hours. ACADL-pEGFP-N1 plasmid was constructed, while mixed RAW264.7 stable clones overexpressing ACADL were selected in neomycin and identified by Western blotting. After 4 hours of LPS stimulation, the secretion of TNF-α, IL-6 and IL-10 were detected by ELISA and their mRNA expression were detected by qRT-PCR. Data showed that in the process of macrophage polarization, the expression of ACADL protein decreased at first and then increased. ACADL overexpression led to secretion increase of IL-6 and IL-10, but not TNF-α, by macrophages in response to LPS. And the mRNA levels of these cytokines showed a similar trend. Beta oxidation inhibitor etomoxir could reduce the secretion of inflammatory factors by RAW264.7 cells. Therefore, ACADL can enhance the expression of IL-6 and IL-10 in macrophages and the mechanism may related to beta oxidation.
作者
黄靖茹
吴宪
邹滔
王庆阳
林周
牛春晓
李淑莲
HUANG Jingru;WU Xian;ZOU Tao;WANG Qingyang;LIN Zhou;NIU Chunxiao;LI Shulian(School of Basic Medicine,Henan University Laboratory of Neuroimmunity and Antibody Engineering,Kaifeng 475000,China;Institute of Military Cognition and Brain Sciences,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2020年第3期219-223,228,共6页
Immunological Journal
基金
国家自然科学基金(31671482)
