PYNOD对LPS活化的BV2小胶质细胞炎症因子释放的影响

Anti-inflammatory effects of PYNOD on lipopolysaccharide-stimulated BV2 microglial cells

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.