›› 2009, Vol. 52 ›› Issue (3): 267-273.doi:

• 研究论文 • 上一篇    下一篇


李有志, 杨海君, 徐汉虹, 刘仲华, 高必达, 张瑞峰, 周志成   

  • 出版日期:2009-04-14 发布日期:2009-03-20
  • 通讯作者: 高必达

Effect of 12a-hydroxyrotenone on the fecundity of Prodenia litura (Fabricius) (Lepidoptera: Noctuidae) and its mechanisms

  • Online:2009-04-14 Published:2009-03-20

摘要: 为探讨12a-羟基鱼藤酮对斜纹夜蛾Prodenia litura (Fabricius)生殖力的影响及其作用机理, 本文测定了该虫取食12a-羟基鱼藤酮后的产卵量。在用高效液相色谱仪确定其卵巢组织中存在12a-羟基鱼藤酮后, 通过MTT法测定该化合物对斜纹夜蛾卵巢细胞(PL细胞)的细胞毒性,并利用流式细胞仪检测该化合物对PL细胞细胞周期、膜电位、胞内钙离子浓度、线粒体膜电位的影响。结果表明:斜纹夜蛾幼虫取食12a-羟基鱼藤酮后产卵量下降80%以上, 并从卵巢组织中检测出该化合物。12a-羟基鱼藤酮对PL细胞增殖具有抑制活性, 其IC50为6.6 mg/L。该化合物将PL细胞周期阻滞于S期, 导致PL细胞膜电位、线粒体膜电位和细胞内游离钙离子浓度均显著升高。该化合物导致斜纹夜蛾繁殖力降低。12a-羟基鱼藤酮将PL细胞增殖阻滞于细胞周期的S期。同时, 该化合物对卵巢细胞具有较弱的毒杀活性, 导致一些卵巢细胞死亡。由于上述原因, 卵巢细胞数量逐渐减少而导致卵巢管萎缩。卵巢管的萎缩使卵巢管内的卵母细胞不能发育成卵细胞, 故其生殖力下降。

关键词: 斜纹夜蛾, 12a-羟基鱼藤酮, 繁殖力, 细胞毒性, 细胞周期, 钙离子浓度, 线粒体膜电位

Abstract: In order to study the effect of 12a-hydroxyrotenone on the fecundity of Prodenia litura (Fabricius) and its action mechanisms, the number of eggs laid per female adult of P. litura was investigated; after 12a-hhydroxyrotenone in ovary was determined with HLPC, its cytotoxicity to ovarian cells from P. litura(PL cells) was determined at 48 h after treatment by MTT assay. The effect of 12a-hydroxyrotenone on cell cycle, cell membrane potential and mitochondrial membrane potential of PL cells, and intracellular free calcium concentration in PL cells at 24 h after treatment was also tested by flow cytometry. The results showed that the number of eggs laid per female adult declined by above 80% after 12a-hydroxyrotenone was fed to P. litura larvae, and the compound could be detected from the ovarian tissues. 12a-hydroxyrotenone showed inhibitory activity to PL cells, with IC50being 6.6 mg/L at 48 h after exposure. After PL cells were treated with 12a-hydroxyrotenone for 24 h, the percentage of S phase cells markedly increased in cultured PL cells (P<0.05), while the percentage of G2-M phase cells decreased. Cell membrane potential, mitochondrial membrane potential and the intracellular free calcium concentration in PL cells were significantly higher than those of the control groups. 12a-hydroxyrotenone led to a linear decrease in the number of eggs laid per female and PL cell cycle arrest in S phase. 12a-hydroxyrotenone exhibited weak cytotoxicity to PL cells, and caused some of them to death. Thus the number of ovarian cells decreased, and the ovary development was inhibited so that ovarian cells could not develop into mature ooctyes and its fecundity obviously decreased.

Key words: Prodenia litura, 12a-hydroxyrotenone, fecundity, cytotoxicity, cell cycle, intracellular free calcium concentration, mitochondrial membrane potential