电化学微传感器实时检测大肠埃希菌激活的RAW264.7细胞释放NO的研究

Study on application of electrochemical microsensor detecting NO released from macrophage stimulated by Escherichia coli

目的探讨NO电化学微传感器在抗细菌感染免疫研究中的应用价值。方法应用前期制备的基于纳米金(nanoAu)修饰玻璃纤维的新型NO电化学微传感器实时检测大肠埃希菌(E.coli)不同浓度刺激组、不同时间刺激组的小鼠巨噬细胞(RAW264.7细胞)NO的释放水平。结果与对照组相比,RAW264.7细胞受到E.coli刺激后,NO的释放水平明显上调(P<0.01),且对E.coli的刺激具有浓度依耐性。随着E.coli作用时间的延续,RAW264.7细胞的NO释放水平逐渐上升,作用12h时达到高峰,然后开始下降。结论 NO电化学微传感器成功应用于E.coli激活的巨噬细胞释放NO过程的实时检测。

Objective To apply nitric oxide（NO） electrochemical microsensor in the real time detection of NO released from RAW 264 .3 cells infected by E .coli ,and to explore the application value of this NO microsensor in the research area of infection im‐munity against bacterium .Methods Taking NO microsensor to detect NO released from RAW 264 .3 cells respectively stimulated by E .coli of different densities and of 1 ×107 mL -1 for different time .Results The level of NO released from RAW 264 .3 cells was enhanced obviously when incubated with E .coli as compared with that of normal cells and the extent of incersase depended on the density of E .coli （P〈0 .01） .The released level of NO increased gradually from the beginning and reached its peal at the time of 12 h then decreased slowly when incubated with E .coli of 1 ×107 mL -1 .Conclusion The electrochemical microsensor was applied in the real time detection of NO released from macrophages activated by E .coli successfully .