基于斑马鱼模型和代谢组学技术筛选天麻中潜在抗癫痫活性成分

doi:10.3976/j.issn.1002-4026.20230089

摘要/Abstract

摘要：

从确有疗效的中药天麻中挖掘潜在的抗癫痫活性成分对于癫痫的防治意义重大。使用斑马鱼幼鱼癫痫模型评价两个产地天麻样本的抗癫痫活性;利用基于LC-MS/QE plus(液相色谱-质谱)的代谢组学技术检测天麻成分,结合模式判别分析方法寻找重要差异代谢物;通过一级质谱与二级质谱数据及文献进行代谢物鉴定,并利用斑马鱼癫痫模型进一步对代谢物的抗癫痫活性进行确认。结果显示9个天麻提取物能够显著减少斑马鱼的涡旋数量,并且两产地天麻在抗癫痫活性上有着显著差异(P<0.05)。代谢组学检测以及模式判别分析筛选并鉴定出了6个重要差异代谢物(巴利森苷E、天麻素、巴利森苷C、巴利森苷D、N6-对羟基苄基腺苷、4, 4'-二羟基二苄基醚)。活性验证发现巴利森苷E、天麻素、N6-对羟基苄基腺苷能够显著抑制斑马鱼癫痫样行为。该研究运用斑马鱼模型与代谢组学找出并确定了天麻中多个具有抗癫痫活性的成分,其中巴利森苷E以及N6-对羟基苄基腺苷首次被发现具有抗癫痫活性,为天麻抗癫痫作用的研究提供了参考价值。

关键词: 天麻, 癫痫, 代谢组学, 斑马鱼, 活性成分

Abstract:

Identifying potential antiepileptic active ingredients in Rhizoma Gastrodiae is of immense significance for the prevention and treatment of epilepsy. In this study, we used a larval zebrafish epilepsy model to evaluate the antiepileptic activity of Rhizoma Gastrodiae from two different regions. In addition, we employed metabolomics technology based on liquid chromatography tandem mass spectrometry/QE plus to detect the ingredients in Rhizoma Gastrodiae and performed pattern analysis to identify key differential metabolites. The metabolites were further identified using primary and secondary mass spectrometry data and literature references. This step was followed by the confirmation of their antiepileptic activity using the zebrafish epilepsy model. The results showed that nine extracts of Rhizoma Gastrodiae significantly reduced the number of whirls in zebrafish, and significant differences in antiepileptic activity were observed between the Rhizoma Gastrodiae samples from the two regions (P<0.05). Metabolomics and pattern analysis identified six important differential metabolites (Parishin E, Gastrodin, Parishin C, Parishin D, N6-p-hydroxybenzyl adenosine, and 4,4'-dihydroxydibenzyl ether). Activity verification results showed that Parishin E, Gastrodin, and N6-p-hydroxybenzyl adenosine significantly inhibited zebrafish epilepsy-like behavior. This study utilized the zebrafish model and metabolomics to identify and determine several active antiepileptic ingredients in Rhizoma Gastrodiae. Of these, Parishin E and N6- p-hydroxybenzyl adenosine were reported for the first time to exhibit antiepileptic activity, thereby serving as a valuable reference for further research on the antiepileptic effects of Rhizoma Gastrodiae.

Key words: Rhizoma Gastrodiae, epilepsy, metabolomics, zebrafish, active ingredients

中图分类号:

R917

陈善军, 王欢, 胡凯庆, 毕文杰, 程贵东, 王松松, 韩利文, 王晓静. 基于斑马鱼模型和代谢组学技术筛选天麻中潜在抗癫痫活性成分[J]. 山东科学, 2024, 37(3): 1-9.

CHEN Shanjun, WANG Huan, HU Kaiqing, BI Wenjie, CHENG Guidong, WANG Songsong, HAN Liwen, WANG Xiaojing. Screening of potential antiepileptic active ingredients in Rhizoma Gastrodiae based on zebrafish model and metabolomics technology[J]. Shandong Science, 2024, 37(3): 1-9.

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中文名称	英文简称	化学式	Δ	保留时间/min	子离子碎片
巴利森苷E	PE	C₁₉H₂₄O₁₃	0.653 4	3.66	173.008 8,111.008 5,
天麻素	GAS	C₁₃H₁₈O₇	0.906 1	1.76	123.045 3,285.099 2,161.045 4,129.978 5
巴利森苷C	PC	C₃₂H₄₀O₁₉	0.096 3	4.20	423.093 5,397.114 2,369.119 1,459.113 9
巴利森苷D	PD	C₂₀H₂₀O₉	1.030 2	6.21	107.049 72,215.016 68
N6-对羟基苄基腺苷	NHBA	C₁₇H₁₉N₅O₅	2.325 3	4.37	136.062 1,242.104 0
4, 4'-二羟基二苄基醚	PHBE	C₁₄H₁₄O₃	0.072 7	6.41	121.029 3,107.050 0,93.034 3