School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, 215123, PR China.
School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, 215123, PR China; Sericulture Institute of Soochow University, Suzhou, Jiangsu, 215123, PR China.
Ecotoxicol Environ Saf. 2019 Jul 30;176:58-63. doi: 10.1016/j.ecoenv.2019.03.074. Epub 2019 Mar 25.
Chlorantraniliprole (CAP) can induce excessive calcium release from muscle of insects, causing muscle paralysis until death, and its residues in farmland can cause poisoning in Bombyx mori (B. mori), resulting in the failure of cocooning. No reports have investigated the effects of CAP exposure on detoxification enzyme activities and detoxification-related gene expression in B. mori. In the present study, we treated mulberry leaves with CAP by the leaf-dipping method, and then B. mori larvae were continuously fed with the polluted mulberry leaves. Moreover, the detoxification enzyme activities and the expressions of detoxification-related genes in the fat body of B. mori were examined. The results showed that at 24 h after CAP exposure, the activities of P450 and GST enzymes were all significantly increased, with P450 enzymes responding fastest. CarE enzyme activity was up-regulated in 24 h, and then it was decreased compared with the control group. Furthermore, the expressions of the key genes in the PI3K/Akt/CncC signaling pathway (PI3K, PDK, Akt, CncC and Keap1) at the mRNA were significantly increased. Western blotting analysis revealed that Akt was inhibited at the protein level, resulting in decreased expression of Keap1 and increased expression of CncC. These results indicated that the PI3K/Akt/CncC signaling pathway in the fat body of B. mori responded to CAP exposure and regulated the expressions of downstream detoxification enzymes, thus enhancing the detoxifying capability of B. mori.
氯虫苯甲酰胺(CAP)可诱导昆虫肌肉过度释放钙离子,导致肌肉麻痹直至死亡,其在农田中的残留会导致家蚕(Bombyx mori,B. mori)中毒,从而导致结茧失败。目前尚无报道研究 CAP 暴露对家蚕解毒酶活性和解毒相关基因表达的影响。在本研究中,我们采用叶浸渍法在桑叶上喷洒 CAP,然后让家蚕幼虫持续食用受污染的桑叶。此外,还检测了家蚕脂肪体中的解毒酶活性和解毒相关基因的表达。结果表明,在 CAP 暴露 24 小时后,P450 和 GST 酶的活性均显著升高,其中 P450 酶的响应最快。CarE 酶活性在 24 小时内上调,然后与对照组相比降低。此外,PI3K/Akt/CncC 信号通路(PI3K、PDK、Akt、CncC 和 Keap1)关键基因的 mRNA 表达显著增加。Western blot 分析显示,Akt 在蛋白水平受到抑制,导致 Keap1 表达降低和 CncC 表达增加。这些结果表明,PI3K/Akt/CncC 信号通路在家蚕脂肪体中对 CAP 暴露做出反应,调节下游解毒酶的表达,从而增强了家蚕的解毒能力。