| 基于GC-MS代谢组学解析阿苯达唑治疗家蚕微粒子病的作用机制 |
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| 引用本文： | 邢东旭,廖森泰,黄文洁,李庆荣,肖阳,赵超艺,晏石娟,蒋满贵,黄旭华,杨琼. 基于GC-MS代谢组学解析阿苯达唑治疗家蚕微粒子病的作用机制[J]. 南方农业学报, 2021, 52(7): 1735-1744. DOI: 10.3969/j.issn.2095-1191.2021.07.001 |
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| 作者姓名： | 邢东旭 廖森泰 黄文洁 李庆荣 肖阳 赵超艺 晏石娟 蒋满贵 黄旭华 杨琼 |
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| 作者单位： | 广东省农业科学院蚕业与农产品加工研究所,广州 510610;广东省农业科学院农业生物基因研究中心,广州 510640;广西蚕业科学研究院,南宁 530007 |
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| 基金项目： | 国家自然科学基金青年基金项目（31702189）；广东省自然科学基金面上项目（2021A1515010595）；广东省农业科技创新及推广体系建设项目（2021KJ124） |
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| 摘 要： | 【目的】利用GC-MS代谢组学技术研究阿苯达唑对患微粒子病家蚕血淋巴代谢物的影响，从代谢组学角度阐明阿苯达唑的作用机制，为以阿苯达唑为主剂研发新的家蚕微粒子病治疗药物提供理论依据。【方法】对五龄起蚕接种家蚕微孢子虫（Nosema bombycis，N.b）建立家蚕微粒子病模型，分别于攻毒后12、24、48、72和96 h开始饲喂阿苯达唑混悬液处理桑叶直至上蔟结茧，以未攻毒未给药的家蚕为对照，待化蛹后逐头镜检调查家蚕感染率，以评价阿苯达唑的治疗效果；同时采用GC-MS代谢组学技术进行非靶向代谢组学研究，筛选患微粒子病蚕体血淋巴中的差异代谢物，并通过MetaboAnalyst 4.0进行代谢通路分析。【结果】阿苯达唑对家蚕微粒子病具有显著的治疗效果，药物作用的关键时间是攻毒后24~48 h。与对照组家蚕血淋巴相比，从模型组家蚕血淋巴中共筛选并定性获得47种差异代谢物，其中27种代谢物呈下降趋势、20种代谢物呈上升趋势。对模型组与阿苯达唑给药组的家蚕血淋巴样本进行比较分析，结果发现除木酮糖、D-葡萄糖-6-磷酸、肌醇、泛酸、甲基丁二酸和油酸外，阿苯达唑对多数与家蚕微粒子病相关的代谢物具有干预调节作用。通过MetaboAnalyst 4.0进行通路分析，发现家蚕感染N.b后有6条主要的代谢通路发生明显变化，分别为:①淀粉和蔗糖代谢；②苯丙氨酸、酪氨酸和色氨酸的生物合成；③苯丙氨酸代谢；④甘氨酸、丝氨酸和苏氨酸代谢；⑤谷胱甘肽代谢；⑥磷酸肌醇代谢。家蚕添食阿苯达唑混悬液处理桑叶后能有效减轻上述代谢通路的改变，从而促使患微粒子病家蚕处于较正常的生理状态。【结论】阿苯达唑对家蚕微粒子病具有显著的治疗效果，药物作用的关键时间在N.b感染后24~48 h，结合N.b的生活史，可全面揭示阿苯达唑治疗家蚕微粒子病的作用机制，即阿苯达唑通过抑制N.b在蚕体内的裂殖体增殖，有效降低N.b感染对家蚕氨基酸代谢和能量代谢的破坏作用，维持家蚕的正常生理状态，而达到治疗效果。
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| 关 键 词： | 家蚕微粒子病 家蚕微孢子虫(N.b) 阿苯达唑 代谢组学 GC-MS |
| 收稿时间： | 2021-05-10 |
Therapeutic mechanism of albendazole in treating pebrine disease of Bombyx mori based on GC-MS metabonomics analysis |
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| XING Dong-xu,LIAO Sen-tai,HUANG Wen-jie,LI Qing-rong,XIAO Yang,ZHAO Chao-yi,YAN Shi-juan,JIANG Man-gui,HUANG Xu-hua,YANG Qiong. Therapeutic mechanism of albendazole in treating pebrine disease of Bombyx mori based on GC-MS metabonomics analysis[J]. Journal of Southern Agriculture, 2021, 52(7): 1735-1744. DOI: 10.3969/j.issn.2095-1191.2021.07.001 |
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| Authors: | XING Dong-xu LIAO Sen-tai HUANG Wen-jie LI Qing-rong XIAO Yang ZHAO Chao-yi YAN Shi-juan JIANG Man-gui HUANG Xu-hua YANG Qiong |
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| Affiliation: | 1 Sericulture and Agri-food Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510610, China;2 Agro-biological Gene Research Center, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;3 Guangxi Academy of Sericulture Sciences, Nanning 530007, China |
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| Abstract: | 【Objective】 GC-MS metabolomics was used to investigate the effect of albendazole on hemolymph metabolites of Bombyx mori with pebrine disease, and to clarify the action mechanism of albendazole from the perspective of metabolomics, so as to provide a theoretical basis for developing new therapeutic drugs for pebrine disease with albendazole as the main agent.【Method】 The model of pebrine disease was established by inoculating 5th instar Bombyx mori larvae with Nosema bombycis(N.b). The mulberry leaves coating with albendazole suspension were fed at 12, 24, 48, 72 and 96 h after inoculation until cocooning. The silkworms without inoculation and administration was used as the control. The infection rate was investigated by individual microscopic examination at pupal stage and the therapeutic effect was evaluated;GC-MS was used to conduct the non-targeted metabonomics study to search the hemolymph related differential metabolites in silkworms with pebrine disease, and the related metabolic pathways were constructed by MetaboAnalyst 4.0.【Result】 The results showed that albendazole had a significant therapeutic effect on pebrine disease, and the key time was 24-48 h after inoculation. Compared with the control group, a total of 47 different metabolites were screened and identified from the hemolymph of B. mori in the model group, of which 27 metabolites showed a downward trend and 20 metabolites showed an upward trend. The comparative analysis of the hemolymph samples between the model group and the albendazole treatment group showed that albendazole had an intervention and regulation effect on most metabolites related to pebrine disease except xylulose, D-glucose 6-phosphate, inositol, pantothenic acid, methylsuccinic acid and oleic acid.Six metabolic pathways changed greatly after infection with N.b by MetaboAnalyst 4.0, which were as follows:① starch and sucrose metabolism;② biosynthesis of phenylalanine, tyrosine and tryptophan;③ phenylalanine metabolism;④ glycine, serine and threonine metabolism;⑤ glutathione metabolism;⑥ inositol phosphate metabolism. The albendazole suspension treatment could effectively reduce the changes of the above metabolic pathways, so as to promote the silkworms suffering from pebrine disease in a more normal physiological state.【Conclusion】 Albendazole had a significant therapeutic effect on pebrine disease of B. mori. The key time of drug action was 24-48 h after N.b infection. Combined with the life cycle of N.b, the mechanism of albendazole in the treatment of pebrine disease can be fully revealed. Albendazole can effectively reduce the the destruction of amino acid metabolism and energy metabolism by N.b infection by inhibiting the proliferation of the meronts in silkworms, thus maintaining the normal physiological state of B. mori and achieve healing effects. |
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