印楝素诱导细胞凋亡途径的研究

徐 霖, 李 升, 王 嘉福
贵阳学院生物与环境工程学院 贵州大学农业生物工程省重点实验室

摘要

目的：印楝素是一种高效低毒的植物源农药，其作用机制目前仍未清楚，深入研究印楝素诱导细胞凋亡的机制有助于进一步利用印楝素农药；方法：采用基因芯片技术结合实时定量 PCR 技术研究印楝素作用过程中相关基因表达差异变化，流式细胞仪结合 Fluo-3 AM 荧光探针检测 Ca2+ 浓度变化；结果：利用基因芯片技术检测到印楝素处理后，与凋亡途径相关 22个基因差异化表达，其中，钙调蛋白（CaM）基因上调 5.05 倍，是上调最多的基因；利用实时定量 PCR 技术验证了凋亡途径中相关基因，证实印楝素通过触发 Ca2+ 释放诱导凋亡；采用流式细胞仪结合 Fluo-3 AM 荧光探针检测到当 Ca2+ 途径受阻，细胞通过蜕皮激素受体途径诱导凋亡；结论：印楝素通过触发 Ca2+ 释放诱导凋亡，并发现 Ca2+ 通过 Ca2+-CaM 信号途径或蜕皮激素受体 ECR/ USP 级联信号途径诱导凋亡。

关键词

印楝素；实时定量 PCR；Ca2+-CaM 信号途径；蜕皮激素受体；ECR/ USP 级联信号途径

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