在体外测试中记者基因的规范化如何影响结果?
Nina Cedergreen1, Geeta Mandava2, Johan Lundqvist2
1Dept. of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark.
Integrated environmental assessment and management
|January 18, 2026
概括
记者基因分析需要仔细的正常化,以准确测量化学生物活性. 使用基线和阳性对照的双重规范化提高了复杂环境样本测试中的可重现性.
科学领域:
- 环境化学环境化学
- 毒理学 毒理学 毒理学
- 生物技术是生物技术.
背景情况:
- 记者基因测试对于评估化学生物活性至关重要.
- 规范化策略显著影响测试解释,但被低估.
- 由于复杂的矩阵和溶剂效应,环境样本存在独特的挑战.
研究的目的:
- 系统地评估不同规范化策略对记者基因检测结果的影响.
- 将三种关键受体系统的正常化方法进行比较:亚利碳化合物受体 (AhR),雌激素受体激动剂 (ERago) 和核因子红色素2相关因子2 (Nrf2).
- 用废水样本评估溶剂暴露和蒸发对测试结果的影响.
主要方法:
- 分析了400多个废水样本,分布在34-54个微板上.
- 规范化方法的比较:减去与根据基线对照进行除法,并对正控进行规范化.
- 评估参考化学物质度-反应曲线,溶剂对照 (甲醇,乙醇),蒸发溶剂和介质对照.
主要成果:
- 低度的溶剂在体外试验中显著改变了受体反应.
- 溶剂蒸发减轻了一些不良影响,但引入了潜在的偏差.
- 测试信号的变化主要是由系统因素 (例如细胞生长,读者灵敏度) 驱动的.
- 对基线和阳性对照的规范化始终改善了EC值的可重现性.
结论:
- 严格的控制验证和规范化对于强大的体外生物测试至关重要,特别是对于环境样本.
- 建议尽量减少溶剂度,验证溶剂效应,并采用双重规范化策略.
- 需要对时间依赖性毒性和矩阵效应进行进一步的研究,以提高试验的可比性.
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