鸦胆因D通过影响胰岛素信号通路和诱导中肠细胞凋亡抑制黑腹果蝇生长  

Bruceine D inhibiting growth of Drosophila melanogaster by influencing insulin signaling pathway and inducing cell apoptosis in midgut

作　　者：陈冬平 李康[2] 龙诗慧 田永清 林菲 徐汉虹[1] CHEN Dongping;LI Kang;LONG Shihui;TIAN Yongqing;LIN Fei;XU Hanhong(College of Plant Protection,South China Agricultural University,Guangdong 510000,China;School of Life Science,South China Normal University,Guangdong 510000,China)

机构地区：[1]华南农业大学植物保护学院,广东广州510642 [2]华南师范大学生命科学学院,广东广州510631

出　　处：《江西农业大学学报》2024年第3期648-655,共8页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(31972286)。

摘　　要：【目的】鸦胆因D(BD)是从中药鸦胆子(Brucea javanica)中提取的植物源农药,具有良好的杀虫活性,能有效抑制包括黑腹果蝇在内的多种害虫的生长发育,但是关于BD的毒性机制研究较少,研究旨在探索BD抑制黑腹果蝇生长的毒性机制。【方法】以黑腹果蝇为研究对象,给2龄末黑腹果蝇喂食含20μg/g BD的人工饲料后,测定用药处理24 h和48 h时,BD对黑腹果蝇体质量、中肠和脂肪体的影响,通过免疫组化、蛋白质免疫印迹和实时荧光定量PCR试验,研究BD是否诱导果蝇中肠细胞凋亡、影响脂肪体细胞生长和影响营养信号通路。【结果】20μg/g BD能显著抑制黑腹果蝇的生长,体现在抑制其体质量增加和脂肪体细胞生长,同时,20μg/g BD处理48 h后,会导致果蝇脂肪体细胞核皱缩和中肠细胞凋亡。实时荧光定量PCR和蛋白质免疫印迹试验发现,BD显著上调胰岛素信号通路负调控基因4EBP和InR的转录水平,同时下调脂肪体4EBP的磷酸化水平,抑制细胞生长。【结论】BD主要通过诱导细胞凋亡和影响胰岛素信号,进而影响营养物质的消化和吸收,最终抑制黑腹果蝇的生长发育。[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 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 20μg/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 20μg/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.

关 键 词：鸦胆因D 抑制生长 凋亡 4EBP INR 

分 类 号：S433[农业科学—农业昆虫与害虫防治] S481.1[农业科学—植物保护]