印楝素对小菜蛾胚胎细胞增殖和凋亡的影响
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国家自然科学基金(31471793,31601699)


Effects of azadirachtin on proliferation and apoptosis of Plutella xyllostella embryonic cells
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    【目的】探讨印楝素对小菜蛾Plutella xyllostella胚胎细胞增殖和凋亡的影响及可能的机制。【方法】将体外培养的小菜蛾胚胎细胞分为印楝素处理组和未处理组,采用CCK-8试剂盒检测细胞增殖的抑制率,激光共聚焦镜检PI染色后的细胞死亡和DAPI染色后的凋亡小体,通过Western-blotting检测各组细胞Caspase-3的表达情况以及通路蛋白Akt的磷酸化水平。【结果】印楝素对小菜蛾胚胎细胞有明显的增殖抑制作用,且呈现浓度依赖性,24 h的IC50为4.4 μg·mL-1。印楝素处理后的细胞经PI染色后能明显观察到死亡细胞,DAPI染色后可见凋亡小体; Caspase-3蛋白发生剪切,并且抑制Akt的磷酸化水平。【结论】印楝素对小菜蛾胚胎细胞的增殖有明显的抑制作用,并通过抑制Akt信号通路的活化,诱导细胞产生依赖于Caspase-3的Ⅰ型凋亡。

    Abstract:

    【Objective】To investigate the effect and mechanism of azadirachtin on proliferation and apoptosis in Plutella xyllostella embryonic cells. 【Method】P. xyllostella embryonic cells were divided into azadirachtin treated and untreated groups, the inhibition rate of cells proliferation was detected using CCK-8 kit. The cell death and apoptotic body were observed using laser confocal microscope after stained by PI and DAPI respectively. The expression of Caspase-3 and the phosphorylation of pathway protein Akt were detected using Western-blotting.【Result】Azadirachtin had significant inhibitory effect on the proliferation of P. xyllostella embryonic cells, and the effect was dose-dependent. IC50was 4.4 μg·mL-1 in 24 h. Dead cells could be observed in azadirachtin treated group after stained by PI, and apoptotic body could be found in the same group after stained by DAPI. Caspase-3 protein was cleaved and Akt phosphorylation was inhibited.【Conclusion】 Azadirachtin has significant inhibitory effect on the proliferation of P. xyllostella embryonic cells. By inhibiting the activation of Akt signaling pathway, azadirachtin can induce cells to produce Caspase-3-dependent type I apoptosis.

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邵雪花,赖多,毛根林,徐汉虹.印楝素对小菜蛾胚胎细胞增殖和凋亡的影响[J].华南农业大学学报,2017,38(4):19-24

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  • 收稿日期:2017-02-21
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  • 在线发布日期: 2017-07-24