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目的:研究神经生长因子(nerve growth factor,NGF)对脊髓C7-T5背根节感觉神经元细胞内诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和P物质表达的影响及干扰素调节因子-1(interferonregulatory factor-1,IRF-1)的调控作用,阐明气道神经源性炎症的机制。方法:体外原代培养C7-T5脊髓背根神经节感觉神经元(DRGn)细胞,然后给予NGF或NGF+IFN-γ刺激。采用实时荧光定量PCR法检测经不同处理后细胞内P物质和iNOS的mRNA表达水平;再采用慢病毒GFP-IRF-1-shRNA转染DRGn细胞,观察阻断IRF-1表达后iNOS和P物质表达的变化。结果:与对照组相比,经NGF刺激后的各组细胞内P物质及iN-OSmRNA水平明显升高(P<0.01);与单纯NGF刺激组相比,NGF+IFN-γ刺激组细胞内P物质mRNA水平无明显变化,而iNOSmRNA水平明显升高(P<0.01);经慢病毒GFP-IRF-1-vshRNA阻断IRF-1表达后,细胞内P物质及iNOS的表达水平明显降低(P<0.01)。结论:NGF通过IRF-1调控气道感觉神经元细胞内P物质及iNOS的合成参与气道神源性炎症,IRF-1可能成为气道神经源性炎症的治疗靶点。
Objective: To investigate the effect of nerve growth factor (NGF) on the expression of inducible nitric oxide synthase (iNOS) and substance P in the sensory neurons of C7-T5 dorsal root ganglion and the effect of interferon Regulatory factor-1 (IRF-1) regulation, clarify the mechanism of airway neurogenic inflammation. Methods: C7-T5 spinal dorsal root ganglion neurons (DRGn) cells were primary cultured in vitro and then stimulated with NGF or NGF + IFN-γ. Real-time fluorescence quantitative PCR was used to detect the mRNA expression levels of substance P and iNOS after treated with different concentrations of GFP-IRF-1-shRNA. The expression of iNOS and substance P Changes in expression. Results: Compared with the control group, the levels of substance P and iN-OS mRNA in each group were significantly increased after NGF stimulation (P <0.01). Compared with the NGF group, NGF + IFN-γ stimulation group (P <0.01). After the expression of IRF-1 was blocked by lentivirus GFP-IRF-1-vshRNA, the expression of substance P and iNOS in cells was significantly decreased (P <0.01) P <0.01). CONCLUSION: NGF participates in airway neurogenic inflammation through regulating the synthesis of substance P and iNOS in airway sensory neurons via IRF-1. IRF-1 may be the target of treatment for airway neurogenic inflammation.