ROS指标在保健食品抗氧化功能检测中的应用及灵敏性比较

APPLICATION AND SENSITIVE COMPARISION OF ROS INDEX ON ANTIOXIDATION EVALUATION OF HYGIENE FOOD

目的:将ROS指标应用于保健食品抗氧化功能检测中,并进行敏感性比较。方法:SPF级昆明种雄性小鼠,12月龄,按体重随机分为老龄组对照组、大豆异黄酮低、中、高剂量组(分别为0·08g/kgBW、0·17g/kgBW、0·50g/kgBW),每组10只动物。连续灌胃给予受试物42d后,断头处死动物,取肝脏。以胶原酶和胰蛋白酶联合酶解肝组织,制备肝组织单细胞悬液;以流式细胞术测定肝细胞ROS水平;同时制备肝组织匀浆,以分光光度法测定肝细胞MDA含量及SOD、GSH-Px活性。结果:流式仪直方图分析显示,随着受试物给予剂量的增加,各剂量组肝细胞ROS水平均明显降低(P<0·05);低、高剂量组肝细胞内MDA含量明显减少(P<0·05),中、高剂量组GSH-Px活性明显增加(P<0·05),高剂量组SOD活性亦明显增加(P<0·05);其余剂量组各指标差异无统计学意义(P>0·05)。结论:大豆异黄酮通过降低肝细胞自由基水平而起到抗氧化作用,以FCM检测肝细胞ROS水平可更好地对保健食品的抗氧化功能进行筛选与评价。

Objective： To apply ROS index in the antioxidation evaluation of hygiene food, and compare its sensitivity with other induces. Methods： 12 - month KM male mice of SPF were randomly grouped into aging controlled group and low, middle, high soy isoflavones groups （0.00 mg/kg b.w, 0.08 mg/kg b.w, 0.17 mg/kg b.w, 0.50 mg/kg b.w separately）. Each group had ten mice. After being given the sample for 42 days, hepatocytes＇ ROS was measured by FC through staining hepatocytes with DCFH- DA, and MBA content, SOD activity, GSH- Px activity were measured by spectrophotometry. Results： Hitogram analysis of FC showed that hepatocytes＇ ROS level in each dose group decreased significantly （P〈0.05）, MDA content in the low and high dose group decreased （P〈0.05）, the activity of GSH- Px in the middle and high group increased （P〈0.05）, the activity of SOD in the high group increased （P〈0.05）. But the index in the rest groups has no significant change （P〉0.05）. Conclusion： Soy isoflavones had the effect of antioxidation by decreasing free radical level, hepatocytes＇ ROS level measured by FC could better appraise the antioxidation system of hygiene food.