| 贝壳杉烷型二萜类化合物PV006对THP-1源性巨噬细胞M1型极化的调节作用 |
| 投稿时间：2017-04-16 修订日期：2017-07-05 点此下载全文 |
| 引用本文：张加芬,李新宇,宋莎莎,王永芳.贝壳杉烷型二萜类化合物PV006对THP-1源性巨噬细胞M1型极化的调节作用[J].医学研究杂志,2018,47(5):64-69 |
| DOI：
10.11969/j.issn.1673-548X.2018.05.016 |
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| 基金项目:中国医学科学院医学与健康科技创新工程项目（2017-12M-1-011） |
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| 中文摘要:目的 利用体外诱导巨噬细胞向M1型极化的模式，初探贝壳杉烷型二萜类化合物PV006对M1型极化的影响。方法 用佛波酯（PMA）联合脂多糖（LPS）及重组人干扰素-γ（rhIFN-γ）刺激人单核细胞株THP-1细胞，促使其向M1型巨噬细胞分化。通过显微镜观察细胞形态学变化，qRT-PCR法检测诱导后的细胞内IL-1β、TNF-α、IL-6以及IL-8 mRNA表达水平，Western blot法检测核转录因子-κB磷酸化P65蛋白（NF-κB p-P65）和磷酸化信号转导与转录激活物1（p-STAT1）蛋白的表达，确认THP-1细胞向M1型极化，同时检测PV006处理后对上述指标的影响。结果 经PMA联合LPS及rhIFN-γ诱导后，THP-1细胞形态由圆形转变为特征的长梭形或纺锤形，IL-1β、TNF-α、IL-6、IL-8 mRNA表达升高，NF-κB p-P65和p-STAT1蛋白表达增强。不同浓度PV006同步处理后，作M1型极化诱导的THP-1细胞未出现特征性的长梭形或纺锤形，大部分细胞仍为圆形。IL-1β、TNF-α、IL-6、IL-8 mRNA表达水平下调（P<0.05），NF-κB p-P65和p-STAT1蛋白表达减弱。结论 PMA联合LPS及rhIFN-γ可诱导THP-1源性巨噬细胞向M1型极化，PV006有抑制巨噬细胞向M1型分化的作用。 |
| 中文关键词:巨噬细胞M1型极化 磷酸化核转录因子-κB P65 磷酸化信号转导与转录激活物1 |
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| Effects of Kaponane Diterpenoids Compound PV006 on M1 Polarization of THP-1-derived Macrophages |
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| Abstract:Objective To study the effects of kaurane diterpenoids compound PV006 on M1 polarization of macrophages by inducing M1 type differentiation of macrophages in vitro.Methods The human monocyte cell line THP-1 cells were stimulated to differentiate into M1-type macrophages by phorbol 12-myristate13-acetate (PMA) combined with lipopolysaccharide (LPS) and recombinant human interferon-γ(rhIFN-γ). M1-type polarization of THP-1-derived macrophages was confirmed by observing morphological changes of cells with microscope, detecting the mRNA expression of IL-1β, TNF-α, IL-6 and IL-8 by real-time fluorescence-based quantitative PCR (qRT-PCR), as well as analyzing expressions of nuclear transcription factor κB phospho-P65 (NF-κB p-P65) and phospho signal transducer and activator of transcription-1 (p-STAT1) by western blotting. After synchronously treatment with PV006, above parameters of stimulated cells were detected.Results After THP-1 cells were induced by PMA combined with LPS and rhIFN-γ, the shape of cells changed from round to characteristic long fusiform or spindle shape, the expression of IL-1β,TNF-α, IL-6, IL-8 mRNA increased, the expression of NF-κB p-P65 and p-STAT1 protiens enhanced. After treatment with different concentrations of PV006, long fusiform or spindle shape cells did not be observed and most cells remains round in M1 polarization stimulated THP-1 cells. PV006 down-regulated the expression of IL-1β, TNF-α, IL-6, IL-8 mRNA(P<0.05), and attenuated the expression of NF-κB p-P65, p-STAT1 proteins.Conclusion M1-type polarization of macrophages derived from THP-1 cells can be induced by stimulation of PMA combined with LPS and rhIFN-γ. PV006 compound could inhibit the differentiation of macrophages into M1 subtype. |
| keywords:Macrophage M1-type polarization Phospho nuclear transcription factor-κB P65 Phospho signal transducer and activator of transcription-1 |
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