一氧化氮光学生物传感器用于药物活性的考察

目的 研制一氧化氮(NO)吸光型生物传感器,并将之用于考察刺激巨噬细胞产生NO药物的活性.方法 以Al2O3/SiO2为载体,溶胶凝胶法固定细胞色素C(Cyt C),组装传感池,通过紫外分光光度(UV)法考察该传感池在酸化的细胞培养液中对NO的响应性能.测定含有γ-干扰素(IFN-γ)、脂多糖(LPS)、以及以上述溶液为阳性对照的硫酸海藻多糖、香菇多糖、甘露聚糖肽的细胞培养液中NO含量.结论 传感池抗干扰能力强,在酸化的培养液中,NO在20.0 μmol/L～180.0 μmol/L的浓度范围内线性关系良好(r＝0.9959),检测限为5.0 μmol/L,日内精密度(RSD)＝3.5%,日间RSD＝7.9%.三种培养液中的NO含量分别为(12.24±3.39) μmol/L、(8.51±0.19) μmol/L、(8.23±0.26) μmol/L.与传统Griess法比较两种方法差异无统计学意义.结论 该传感器法可以对细胞培养液中的NO进行半定量分析,作为药物活性初筛的一种新手段.

Research on Activities of Drugs by Use of an Optical Nitric Oxide Biosensor

OBJECTIVE: To develop an Optical Nitric Oxide Biosensor and investigate the activities of drugs that stimulate macrophage to produce nitric oxide. METHODS: An NO biosensor was built on Cyt C which had been immobilized by Al2O3/SiO2 sol-gel procedure. Then the biosensor's characteristics in response to nitric oxide were studied by UV. The detection of NO in the culture solution that contained drugs (sulfated polysaccharides, Lentinan, Mannatide) was carried out by using interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) as positive control. RESULTS: The standard curve was linear within the range of 20. 0-180.0 micromol/L of NO in acidified culture solution (r = 0.9959). The limit of detection was 5.0 micromol/L; within-day precision was 3.5%; day-to-day precision was 7.9%. The concentrations of NO in culture solutions were (12.24+/-3.39) micromol/L, (8.51 +/- 0.19) micromol/L and (8.23 +/- 0.26) micromol/L, which were compared with the use of Griess method, and no statistically significant difference was observed. CONCLUSION: The biosensor can be used as a semi-quantitative analysis method and be used for screening drugs preliminarily.