小鼠白细胞介素-10表达载体的构建及其在N9细胞中的表达

Construction of Fluorescent Eukaryotic Expressing Vector of Mouse Interleukin-10 and its Expression in the N9 Cells

  • 摘要: 白细胞介素10(interleukin 10,IL-10)是免疫系统最重要的抗炎症细胞因子之一,具有神经保护作用。为了探究IL-10发挥神经免疫的抗炎作用,该文首先采用反转录-聚合酶链式反应(RT-PCR)技术从小鼠脾脏组织总RNA中扩增出该基因的全序列cDNA;再以腺相关病毒(AAV)为载体,构建了带有高亮度增强型绿色荧光蛋白(EGFP)报告基因的真核表达载体AAV-IL10-EGFP,观察了其在小胶质细胞系N9细胞中的表达状况;最后采用实时荧光定量(QT-PCR)的方法,检测AAV-IL10-EGFP在N9细胞中的抗炎作用。

     

    Abstract: Interleukin-10(IL-10) is an important anti-inflammatory cytokine in the immune system and has neuroprotective effects. To explore the anti-inflammatory effect of IL-10 on neuroimmunomodulatory mechanisms, the full-sequence cDNA of the gene was amplied from the total RNA of mouse spleen by reverse transcription-polymerase chain reaction(RT-PRC). AAV-IL10-EGFP was constructed with adeno-associated virus (AAV) as a vector and a high-intensity enhanced green fluorescent protein (EGFP) reporter gene. The expression of AAV-IL10-EGFP was successfully achieved in N9 microglial cells, which showed inhibitory effects on the production of pro-inflammatory cytokines.

     

/

返回文章
返回