摘要
目的:观察PYNOD对LPS活化的BV2小胶质细胞炎症因子释放的影响。方法:将表达PYNOD的重组质粒p EGFP-C2-PYNOD瞬时转染BV2细胞后,加入LPS作用24 h,Griess法检测一氧化氮(nitric oxide,NO)的释放,实时荧光定量PCR(real-time PCR)检测诱导型一氧化氮合酶(inducible NO synthase,i NOS)和白细胞介素-1β(interleukin-1β,IL-1β)mRNA的表达,此外Western blotting和ELISA法检测i NOS和IL-1β的蛋白表达。结果:转染PYNOD重组质粒能显著抑制LPS诱导的BV2小胶质细胞炎症因子NO的释放(P<0.05)。Real-time PCR证实PYNOD可抑制i NOS和IL-1β的mRNA表达,差异有统计学意义(P<0.05)。ELISA和Western blotting证实PYNOD可下调i NOS和IL-1β蛋白的表达(P<0.05)。结论:PYNOD蛋白可以在转录水平和翻译水平显著抑制LPS刺激的BV2小胶质细胞活化产生的炎症反应。
AIM： To validate the anti-inflammatory effect of PYNOD on lipopolysaccharide（ LPS）-stimulated BV2 microglial cells. METHODS： A specific GFP and PYNOD fluorescence expression vector p EGFP-C2-PYNOD driven by the promoter of CMV gene was constructed. The anti-inflammatory properties of PYNOD were studied using LPS-stimulated BV2 microglia model. The productions of nitric oxide（ NO）,inducible NO synthase（ i NOS）,interleukin-1β（ IL-1β）which caspase-1 were evaluated as inflammatory parameters. RESULTS： Pretreatment with p EGFP-C2-PYNOD to BV2 microglia cells stimulated by LPS significantly inhibited the excessive productions of NO and IL-1β,which was associated with down-regulation of i NOS and caspase-1 at mRNA and protein levels. CONCLUSION： PYNOD might be useful for treating the inflammatory and deleterious effects of BV2 microglial cell activation in response to LPS stimulation.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第11期1954-1959,共6页
Chinese Journal of Pathophysiology
基金
赣州市科技计划项目(赣市财教字[2013]22号)
赣南医学院重点课题(No.2D201201)
