LncRNA-Scarna10对白藜芦醇促进巨噬细胞M2极化影响的实验研究

Experimental study on the effect of lncRNA-Scarna10 on resveratrol-induced M2 polarization of macrophages

  • 摘要:
    目的  探讨lncRNA-Scarna10(Scarna10)对白藜芦醇(RSV)促进巨噬细胞极化的影响。
    方法  LPS构建RAW264.7巨噬细胞M1极化模型,RSV干预24 h,设置分组:control组、LPS组、RSV+LPS组,采用实时荧光定量聚合酶链式反应(RT-qPCR)检测Scarna10表达水平,蛋白质免疫印迹法(western blotting)及免疫荧光检测巨噬细胞极化标志物(iNOS、Arg-1、CD206)表达。在RAW264.7细胞中沉默Scarna10,分为Si-NC组、LPS+Si-NC组、RSV+LPS+Si-NC组、RSV+LPS+Si-Scarna10组,检测转染后各组细胞内Scarna10、巨噬细胞极化标志物的表达。
    结果  经LPS诱导及白藜芦醇干预后,LPS组M1极化标志物iNOS蛋白表达水平升高,RSV+LPS组M2极化标志物Arg-1、CD206蛋白表达水平均升高,iNOS的蛋白表达水平下降,同时Scarna10在RSV+LPS组的表达较LPS组升高;沉默Scarna10后,与RSV+LPS+Si-NC组比较,RSV+LPS+Si-Scarna10组Scarna10表达及Arg-1、CD206蛋白表达水平均下降,iNOS的蛋白表达水平升高。以上各组间的差异均有统计学意义(均P<0.05)。
    结论  Scarna10在RSV干预后的M1型巨噬细胞中表达升高,沉默Scarna10可以逆转RSV对M1型巨噬细胞向M2型转化的促进作用

     

    Abstract:
    Objective  To investigate the effect of lncRNA-Scarna10 (Scarna10) on resveratrol (RSV)-induced polarization of macrophages.
    Methods  M1 polarization model of RAW264.7 macrophages was established by LPS and treated with RSV for 24 hours. The study groups were divided into control group, LPS group and RSV+ LPS group. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of Scarna10, and western blotting and immunofluorescence were used to detect the expression of macrophage polarization markers (iNOS, Arg-1, CD206). RAW264.7 cells were divided into Si-NC group, LPS+Si-NC group, RSV+LPS+Si-NC group and RSV+LPS+Si-Scarna10 group after silencing Scarna10, and the expression of Scarna10 and macrophage polarization markers in each group after transfection was detected.
    Results  After LPS induction and resveratrol intervention, the protein expression level of iNOS, a M1 polarization marker, was elevated in the LPS group, the protein expression levels of Arg-1 and CD206, M2 polarization markers, were elevated, while the protein expression level of iNOS was decreased in the RSV+LPS group. At the same time, the expression level of Scarna10 in the RSV+LPS group was higher than that in the LPS group. After silencing Scarna10, compared with the RSV+LPS+Si-NC group, the expression level of Scarna10 and the protein expression levels of Arg-1 and CD206 in RSV+LPS+Si-Scarna10 group were decreased, and the protein expression level of iNOS was elevated. The differences among the above groups were statistically significant (all P < 0.05).
    Conclusion  The expression of Scarna10 is upregulated in M1 macrophages following RSV intervention, and silencing Scarna10 can reverse the RSV-induced promotion of M1-to-M2 macrophage polarization.

     

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