摘要
目的探讨PPARγ对缺氧复氧RAW264．7小鼠巨噬细胞异质性的影响。方法体外培养小鼠巨噬细胞系（RAW264．7），制备缺氧复氧模型，用慢病毒包装进行PPARγ过表达和基因沉默，细胞分为对照组、模型组、过表达PPARγ组和PPARγ基因沉默组。用ELISA法、RT—PCR、Western印迹法和免疫荧光法检测各组细胞巨噬细胞亚型M1标志物诱导型一氧化氮合酶（iNOS）、M2标志物CD206及肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）的表达。结果与对照组相比，ELISA法检测模型组TNF—α和IFN-γ水平明显升高（均P〈0．01），RT—PCR、Western印迹法和免疫荧光检测均显示TNF-α、IFN-γ、iNOS表达上调，CD206表达下调。与模型组相比，过表达PPARγ湿著减轻了IRI导致的TNF-α、IFN-γ和iNOS表达的上调，上调了CD206表达；与此相反，PPAR1基因沉默加重IRI导致TNF—α、IFN-γ和iNOS表达上调，下调了CD206表达。结论PPARγ在小鼠RAW264．7巨噬细胞缺氧复氧模型中参与调节巨噬细胞表型转换。
Objective To investigate whether peroxisome proliferator activated receptor-γ （ PPARγ ） is involved in oxygen-glucose deprivation （OGD）/reperfusion-induced expression of inflammatory mediators in RAW264.7 murine macrophages and their phenotype switch. Methods OGD/reperfusion-stimulated RAW264.7 murine macrophages served as a model of I/R injury. Knockdown and over expression of PPARγ in RAW264.7 cells were performed. The cells were divided into a control group, a model group, a model ＋PPARγ group, and a model ＋si-PPAR 7 group. The M1 marker inducible nitric oxide synthase （ iNOS ） , M2 marker cluster of differentiation （CD）206, tumor necrosis factor-α （ TNF-α） , and interferon-γ（IFN-γ） were separately examined by ELISA, RT-PCR, Western blot analysis, and immunofluorescence staining. Results The levels of TNF-α and IFN-γ were significantly higher in the model group than in the control group （ bothP 〈 0.01 ） . RT-PCR, Western blot analysis, and immunofluorescence staining showed that the expressions of iNOS, TNF-α, and IFN-γ were increased and that of CD206 decreased. The over-expression of PPAR-γ significantly attenuated OGD/reperfusion- induced alterations and increased the expressions of TNF-α, IFN-γ, and iNOS and down-regulated the expression of CD206. Conversely, the knockdown of PPARγ expression in these cells increased the OGD/reperfusion-induced expressions of iNOS, TNF-α, and IFN-γ and enhanced the OGD/reperfusion-induced downregulation of the expression of CD206. Conclusion PPARγ is involved in macrophage heterogeneity and IR-induced expression of inflammatory mediators in ischemia-reperfusion injury.
出处
《国际医药卫生导报》
2017年第15期2330-2334,2337,共6页
International Medicine and Health Guidance News
基金
广东省科技计划项目（2014A020212519）
广东省中医药强省科研课题（20151060）
