【摘 要】
：
[目的]文献报道黄酮类化合物可通过促进肿瘤坏死因子及抗氧化等途径发挥抗肿瘤作用,我们的前期研究利用Bhas 42细胞两阶段转化实验发现染料木黄酮可能具有非遗传毒性致癌性.本研究采用Bhas 42细胞高通量转化试验对含黄酮类化合物有毒中药芫花和了哥王潜在致癌性进行评价.应用流式细胞仪检测分析芫花、了哥王对Bhas42细胞细胞周期的影响.[方法]1)芫花和了哥王根经10倍体积乙醇(95％)浸提后,制成
【机 构】
：
中国食品药品检定研究院国家药物安全评价监测中心;中国药科大学药学院 中国食品药品检定研究院国家药物
【出 处】
：
中国毒理学会第七次全国毒理学大会暨第八届湖北科技论坛
论文部分内容阅读
[目的]文献报道黄酮类化合物可通过促进肿瘤坏死因子及抗氧化等途径发挥抗肿瘤作用,我们的前期研究利用Bhas 42细胞两阶段转化实验发现染料木黄酮可能具有非遗传毒性致癌性.本研究采用Bhas 42细胞高通量转化试验对含黄酮类化合物有毒中药芫花和了哥王潜在致癌性进行评价.应用流式细胞仪检测分析芫花、了哥王对Bhas42细胞细胞周期的影响.[方法]1)芫花和了哥王根经10倍体积乙醇(95％)浸提后,制成芫花和了哥王粗提干粉.2)Bhas 42细胞高通量转化试验：采用CCK-8法测定两种提取物在启动和促癌阶段对Bhas42细胞相对生长率的影响,确定5个浓度梯度开展基于96孔板的高通量Bhas 42细胞转化试验.接种后第21天,采用H2O2方法判定细胞转化程度.结果用One-way ANOVA(Dunnett-t)检验分析.3)采用流式细胞仪Propidium Iodide (PI)单标法检测芫花了哥王对Bhas42细胞细胞周期的影响,试验设置空白对照组、芫花三个剂量组(0.3、1和3μg·mL-1)以及了哥王三个剂量组(0.3、1和3μg·mL-1).[结果]1)Bhas 42细胞高通量转化试验：在启动试验中,芫花各个浓度组的平均OD值相对阴性对照组(0.27±0.09)均无显著性差异(p＞0.05)；了哥王10 μg·mL-1时结果为阳性(p＜0.05,0.59±0.42)；在促癌试验中,芫花处理浓度为0.3、1、3μg·mL-1时(平均OD值分别为：0.92±0.52、1.40±0.43和1.24±0.41)相对对照组(0.30±0.13)有显著性差异(p＜0.05)；了哥王处理浓度为0.1～10 μg·mL-1时(平均OD值分别为：2.24±0.86、2.89±0.36、2.77±0.36、2.76±0.38和2.83±0.40)相对于阴性对照组均有显著性差异(p＜0.05).2)流式细胞仪检测对Bhas42细胞周期影响：与正常对照组(S期：19.44±3.41％)相比,芫花三个剂量组0.3、1和3μg·mL-1(S期分别占28.02±2.07％、40.85±3.22％、46.18±3.70％)、了哥王三个剂量组0.3、1和3μg·mL-1(S期分别占26.93±1.40％、39.49±1.20％、40.57±4.25％)细胞G1期百分比降低,S期百分比明显升高,并呈剂量相关性.[结论]Bhas 42细胞转化试验可较为灵敏而简便地对化合物非遗传毒性致癌性进行评价.启动试验结果提示芫花和了哥王提取物不具有遗传毒性致癌性,而促癌试验结果提示两者可能具有非遗传毒性致癌性.流式细胞术可以可准确分析单个细胞所处的细胞周期相,定量的反应群体细胞所处的状态,流式细胞术细胞周期分析结果验证了芫花、了哥王对Bhas42细胞增殖转化作用.
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