奇正消痛贴膏提取物对脂多糖诱导的巨噬细胞一氧化氮及其诱导型合酶的影响

Effects of extracts of Cheezheng pain relieving plaster on nitric oxide and iNOS expression in macrophages induced by lipopolysaccharides

考察奇正消痛贴膏提取物(extracts of Cheezheng pain relieving plaster,ECPRP)对巨噬细胞一氧化氮的生成及诱导型一氧化氮合酶表达的影响,并探讨其作用机制。采用Griess试剂检测培养上清液中NO2-的含量,RT-PCR法测定LPS刺激的小鼠腹腔巨噬细胞中iNOSmRNA的表达,Western blotting法测定其iNOS蛋白的表达和NF-κB的活化情况。结果显示,ECPRP(62.5和125mg·L-1)可显著抑制巨噬细胞NO的生成以及LPS引起的iNOS mRNA和蛋白表达的增加;并可抑制I-κB在细胞浆内的降解以及NF-κBp65向胞核内的转位、阻止其活化。因此,ECPRP的抗炎作用机制之一可能是通过抑制巨噬细胞NF-κB的活性,从而降低巨噬细胞iNOSmRNA和蛋白的表达、减少NO的生成。

This study is to explore the effects of extracts of Cheezheng pain relieving plaster （ECPRP） on nitric oxide （NO） production and expression of inducible nitric oxide synthase （iNOS） in macrophages induced by LPS and the mechanism involved. Nitric oxide level was measured with Griess reagent assay. Inducible nitric oxide synthase （iNOS） protein and NF-κBp65 fragment were detected with Western blotting. ECPRP （62.5 and 125 mg·L^-1） significantly inhibited the increase of nitric oxide level. Furthermore, ECPRP （62.5 and 125 mg·L^-1） notably reduced the expression of iNOS mRNA and protein. ECPRP （62.5 and 125 mg·L^-1） elevated the content of I-κB protein in cytoplasm, while decreased the content of NF-κBp65 protein in nucleus. These results suggest that ECPRP reduce nitric oxide level via down-regulation of NF-κB-iNOS-nitric oxide pathway, resulting in prevention of inflammation.