Bruceine D inhibiting growth of Drosophila melanogaster by influencing insulin signaling pathway and inducing cell apoptosis in midgut
[Objective]Bruceine D is a botanical pesticide isolated from the traditional Chinese medicine Brucea javanica(Linn.)Merr.It has good insecticidal activity and can effectively inhibit the growth of various insect pests including Drosophila melanogaster.However,the toxicology mechanism is limited.This study aims to explore the toxic mechanism of BD of inhibiting the growth of D.melanogaster.[Method]Using D.melanogaster as the research object.End 2nd instar larvae of D.melanogaster were treated artificial diet containing 20 μg/g BD.The effects of BD on body weight,midgut and fat body of D.melanogaster after treatment for 24 h and 48 h were studied.We performed experiments of immunocytochemistry,western blot,quantitative real time PCR to investigate whether BD can induce midgut apoptosis,affect the growth of fat body cells and affect nutritional signaling pathway in D.melanogaster.[Result]It was found that 20 μg/g BD can significantly inhibit weight gain and growth of adipose somatic cells of D.melanogaster.Meanwhile,the cell nucleus of the fat body was wrinkled and apoptosis was induced in midguts after 48 h BD treatment.In addition,the experiment of qPCR and western blot showed that BD significantly up-regulated the transcription levels of insulin signaling pathway negative regulatory genes 4EBP and InR of the insulin signaling pathway,while down-regulating the phosphorylation level of 4EBP of fat body,which inhibited cell growth.[Conclusion]These results showed that BD could induce cell apoptosis and suppress insulin signaling,potentially affecting nutrient digestion and absorption,ultimately inhibiting the growth and development of D.melanogaster.
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