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为了明确印楝素A和B活性差异的机理,本研究比较了印楝素A和印楝素B对粉纹夜蛾Trichoplusia ni离体培养胚胎细胞系BTI-Tn-5B1-4的毒性。结果表明:印楝素A与印楝素B对BTI-Tn-5B1-4细胞具有良好的增殖抑制活性,处理后3d,其IC50值分别为2.09μg/mL和9.85μg/mL,印楝素A的细胞毒力显著高于印楝素B。倒置显微镜观察发现,印楝素A和印楝素B处理可导致细胞变形,贴壁能力下降,并出现明显空泡,印楝素A的影响明显高于印楝素B。流式细胞仪检测结果表明,印楝素可导致BTI-Tn-5B1-4细胞体积显著膨大,印楝素A处理细胞体积增大程度显著高于印楝素B;印楝素可以明显影响BTI-Tn-5B1-4细胞膜电位,1.25μg/mL印楝素A和印楝素B处理后3d,细胞DiBAC4(3)荧光强度分别增加88.12%和55.37%,印楝素A的影响显著高于印楝素B。荧光显微镜观察发现,印楝素对BTI-Tn-5B1-4细胞核具有明显影响,印楝素B的影响明显高于印楝素A,印楝素B处理后,细胞核受损细胞数更多,受损程度更严重。结果显示印楝素A和印楝素B的细胞作用机理存在差异,本研究从细胞学水平解释了印楝素的生长发育抑制作用机理。
In order to clarify the mechanism of the difference in the activities of azadirachtin A and B, this study compared the toxicity of azadirachtin A and azadirachtin B to the in vitro cultured embryonic cell line BTI-Tn-5B1-4 of Trichoplusia ni. The results showed that azadirachtin A and azadirachtin B had good proliferation inhibitory activity on BTI-Tn-5B1-4 cells. IC50 values were 2.09μg / mL and 9.85μg / mL respectively after treatment, and azadirachtin The cytotoxicity of A was significantly higher than that of Azadirachtin B. Inverted microscopy showed that azadirachtin A and azadirachtin B resulted in cell deformability and decreased adherence, with obvious vacuoles. The effect of azadirachtin A was significantly higher than that of azadirachtin B. Flow cytometry results showed that azadirachtin could cause significant volume expansion of BTI-Tn-5B1-4 cells, azadirachtin A-treated cells significantly increased the volume of more than azadirachtin B; azadirachtin can significantly affect the BTI The fluorescence intensity of DiBAC4 (3) increased by 88.12% and 55.37% respectively after treated with 1.25μg / mL azadirachtin A and azadirachtin B for 3h-Tn-5B1-4 cell membrane potential. The influence of azadirachtin A was significantly Azadirachtin B. Fluorescence microscopy showed that azadirachtin had a significant effect on the nuclei of BTI-Tn-5B1-4 cells. The effect of azadirachtin B was significantly higher than that of azadirachtin A. After azadirachtin B treatment, Damage is more serious. The results showed that the mechanism of action of azadirachtin A and azadirachtin B were different. In this study, the cytotoxic effect of azadirachtin on the growth and development of azadirachtin was explained.