家蚕幼虫体内抗氧化活性物质成分筛选及非靶向代谢组学分析  

Screening of antioxidant active components from the larval bodies of Bombyx mori and analysis of non-targeted metabolomics

作　　者：谢岩 孙志超 高妍夏 周玲 刘麟菲 王晖 XIE Yan;SUN Zhi-Chao;GAO Yan-Xia;ZHOU Ling;LIU Lin-Fei;WANG Hui(Applied Technology R&D Center for Special Sericulture of Hebei Province Universities,Institute of Sericulture,Chengde Medical University,Chengde 067000,China)

机构地区：[1]承德医学院蚕业研究所,河北省高校特色蚕桑应用技术研发中心,承德067000

出　　处：《昆虫学报》2024年第11期1461-1474,共14页Acta Entomologica Sinica

基　　金：河北省自然科学基金青年项目(C201806033);河北省高等学校科学技术研究项目(BJ2018032);河北省科技厅“技术创新引导专项-科技工作会商”项目;河北省高层次人才资助项目(C20231120);河北省中医药管理局科研计划项目(2022160)。

摘　　要：【目的】分析家蚕Bombyx mori幼虫体内天然主效抗氧化成分的类别、作用效果和单体组成,为其开发利用以及药效作用的研究提供参考。【方法】以2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid),ABTS]和2,2-二苯基-2-三硝基苯肼(2,2-diphenyl-1-picrylhydrazyl,DPPH)自由基的清除能力测定3个桑树品种(禾叶白、冀桑2号和桲椤)叶以及取食冀桑2号叶片的4个家蚕品种(菁松×皓月、东肥、彩3和彩4)5龄幼虫蚕砂和全蚕体的提取液的抗氧化能力(antioxidant capacity,AC);采用分光光度法和反相高效液相色谱-紫外检测法(reverse-phase high-performance liquid chromatography with ultraviolet detection,RP-HPLC-UV)测定全部提取液的总黄酮、总酚、多糖和1-脱氧野尻霉素(1-deoxynojirimycin,1-DNJ)的含量,分析幼虫全蚕体的主效抗氧化活性物质;检测6个家蚕品种(东肥、安四无花、彩甘、7903、彩0和彩3)末龄(3眠蚕的4龄,4眠蚕的5龄)幼虫每日血淋巴中的总酚含量和清除ABTS自由基的AC(AC ABTS);采用液相色谱-质谱联用技术(liquid chromatograph-mass spectrometer,LC-MS)对4个家蚕品种(菁松×皓月、东肥、彩3和彩4)5龄最后1 d幼虫全蚕体和冀桑2号叶片抗氧化活性物质成分进行非靶向定性和定量解析。【结果】桑叶提取液的总黄酮、总酚和多糖含量与提取液的AC呈现极显著正相关,5龄幼虫蚕砂和全蚕体提取液中总黄酮和总酚含量与其提取液的AC呈极显著正相关。4个家蚕品种5龄最后1 d幼虫全蚕体提取液的AC ABTS[(22.61±1.37)~(36.70±2.12)mg VC/g]和AC DPPH[(16.77±0.47)~(23.43±2.34)mg VC/g]分别是冀桑2号叶片提取液的1.46~2.38和1.81~2.54倍,也分别是5龄最后1 d幼虫蚕砂提取液的0.96~2.27和1.23~7.73倍,5龄最后1 d幼虫全蚕体提取液的总酚含量[(9.14±0.13)~(13.28±0.78)mg/g]与冀桑2号叶提取液的[(19.74±0.58)mg/g]和5龄最后1 d幼虫蚕砂的[(10【Aim】To analyze the types,effects and monomer composition of the main natural antioxidant components in the larval body of Bombyx mori,so as to provide a reference for their development and utilization as well as the study of their pharmacodynamic effects.【Methods】The antioxidant capacity(AC)of the extracts from the leaves of three varieties of mulberry trees(Heyebai,Jisang2 and Boluo),as well as the extracts from excrement and whole larval bodies of the 5th instar of four varieties of B.mori(Jingsong×Haoyue,Dongfei,Cai3 and Cai4)fed on Jisang2 leaves,were tested by the scabenging abilities to free radicals of 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)(ABTS)and 2,2-diphenyl-1-picrylhydrazyl(DPPH).Meanwhile,the contents of total flavonoids,total phenols,polysaccharids and 1-deoxynojirimycin(1-DNJ)in the above extracts were determined by spectrophotometry and reverse-phase high-performance liquid chromatography with ultraviolet detection(RP-HPLC-UV)to analyze the main antioxidant active substances from the whole larval body of B.mori.The daily contents of total phenol and the AC for scavenging ABTS radicals(AC ABTS)in the hemolymph of the last instar larvae(the 4th instar of 3-molter silkworms,the 5th instar of 4-molter silkworms)of six varieties of B.mori(Dongfei,Ansiwuhua,Caigan,7903,Cai0 and Cai3)were detected.Furthermore,liquid chromatograph-mass spectrometer(LC-MS)was used for the non-targeted qualitative and quantitative analysis of antioxidant active components in the whole bodies of four varieties of larval B.mori(Jingsong×Haoyue,Dongfei,Cai3 and Cai4)on the last day of the 5th instar and in the leaves of Jisang2.【Results】The contents of total flavonoids,total phenols and polysaccharids of the extracts from mulberry leaves showed highly significant positive correlations with the AC of extracts from mulberry leaves,and the contents of total flavonoids and total phenols showed highly significantly positive correlations with the AC of extracts from excrement and whole bodies of the 5

关 键 词：家蚕 全蚕体 抗氧化能力 酚类化合物 非靶向代谢组 解毒酶基因 

分 类 号：S881[农业科学—特种经济动物饲养]