基于代谢组学的麝鼠香腺上皮细胞中降麝香酮生物合成机制研究  

Biosynthesis mechanism of normuscone in muskrat gland epithelial cells based on metabonomics

作　　者：赵贵军[1,2] 谌颖莲 曾德军 竭航[1] 张承露 吴佳勇[1] 封孝兰[1] ZHAO Gui-jun;CHEN Ying-lian;ZENG De-jun;JIE Hang;ZHANG Cheng-lu;WU Jia-yong;FENG Xiao-lan(Chongqing Institute of Medicinal Plant Cultivation,Nanchuan,Chongqing 408435,China;Chongqing Jinfo Mountain Forest Ecosystem Field Scientific Observation and Research Station,Nanchuan,Chongqing 408435,China)

机构地区：[1]重庆市药物种植研究所,重庆南川408435 [2]重庆金佛山森林生态系统野外科学观测研究站,重庆南川408435

出　　处：《西南农业学报》2022年第10期2462-2468,共7页Southwest China Journal of Agricultural Sciences

基　　金：重庆市自然科学基金项目(cstc2019jcyj-msxmX0024)。

摘　　要：【目的】构建麝鼠香腺上皮细胞的睾酮体外诱导模型,测定分析睾酮诱导培养的香腺上皮细胞代谢物差异表达情况,探究诱导刺激模型下环碳分子架构的生物合成过程,解析麝鼠香腺上皮细胞中降麝香酮生物合成机制。【方法】取9只12~18月龄繁殖期雄性麝鼠,将其香腺组织分离后,通过组织块培养获取离体香腺上皮细胞,使用睾酮诱导培养,以繁殖期麝鼠的香腺和肝组织样本为对照,采用LC-MS非靶向代谢组学技术测定其代谢物表达情况,再对过滤后的高质量数据进行主成分分析、差异代谢物KEGG通路富集分析。【结果】睾酮诱导离体培养香腺上皮细胞48 h后,在TP诱导组细胞培养液中能够检测出大量雄性激素类似物(睾酮、睾酮β-D-葡糖苷酸、甲羟孕酮、前列腺素等)的高表达,以及甲氧麻黄酮、犬尿酸、N-乙酰-L-谷氨酸和黄曲霉素G1类分子的低表达;色氨酸、精氨酸和长链脂肪酸代谢物呈显著高表达,提示睾酮诱导能够促进麝香上皮细胞的脂肪酸碳链和部分氨基酸的生物合成;分析繁殖期麝鼠香腺和肝脏组织中的差异代谢物,结果显示相对于肝脏组织,肌苷、乙酰肉碱、苏氨酸等代谢物在香腺中高表达,进一步提示降麝香酮的生物合成涉及氨基酸和长链脂肪酸代谢。【结论】睾酮诱导离体培养麝鼠香腺上皮细胞后,其色氨酸、精氨酸和长链脂肪酸代谢通路发生显著变化;繁殖期麝鼠香腺组织中肌苷、乙酰肉碱、苏氨酸等代谢物含量较肝脏中高,提示麝鼠中降麝香酮生物合成涉及氨基酸和长链脂肪酸代谢,为解析麝鼠香的生物合成机制提供基础数据和新研究方向。【Objective】The study aimed to construct a testosterone-induced model of muskrat gland epithelial cells in vitro, measure and analyze the differential expression of metabolites in muskrat incense gland epithelial cells induced by testosterone, to explore the biosynthetic process of the cyclic carbon molecular architecture under the induced stimulation model, and analyze the biosynthesis mechanism of normuscone in muskrat fragrant gland epithelial cells.【Method】Nine male muskrats aged 12-18 months in reproductive period were selected, and their fragrance glands were separated, then the epithelial cells of their fragrance glands in vitro were obtained by tissue block culture, which was induced and cultured with testosterone. The fragrance glands and liver tissue samples of muskrats in reproductive period were used as controls, and the expression of metabolites was measured by LC-MS non-targeted metabolomics. Then, the filtered high-quality data were subjected to principal component analysis and differential metabolite KEGG pathway enrichment analysis.【Result】After 48 hours of TP-induced culture of fragrant gland epithelial cells in vitro, high expression of androgen analogs(testosterone, testosterone β-D-glucuronide, medroxyprogesterone, prostaglandins, etc.) could be detected in the TP-induced group, as well as low expression of mephedrone, canine uric acid, N-acetyl-L-glutamic acid and aflatoxin G1 molecular;Tryptophan, arginine and long-chain fatty acid metabolites were significantly overexpressed, suggesting that testosterone induction can promote the biosynthesis of fatty acid carbon chains and some amino acids in musk epithelial cells;Analysis of the differential metabolites in muskrat’s fragrant gland and liver tissue during the breeding period showed that inosine, acetylcarnitine, threonine and other metabolites were highly expressed in fragrant gland compared with liver tissue, further suggesting that the biosynthesis of normuscone involves the metabolism of amino acids and long-chain fatty a

关 键 词：麝鼠 代谢组学 睾酮 降麝香酮 香腺组织 

分 类 号：S865.27[农业科学—野生动物驯养] O657[农业科学—畜牧兽医]