精油香气抗焦虑作用的代谢组学研究
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摘要
焦虑症是目前常见的流行性精神问题之一,在全世界范围内影响着很多人的身心健康。通过嗅闻精油香气或者精油透过皮肤按摩吸收,可以调整及改善人的生理、情绪以及心理状况。虽然临床研究以及芳香疗法应用实践表明精油具有明显的治疗效果,但是有关于精油功效的科学证据依然相当匮乏,尤其缺乏采用严密的分析方法捕捉对人体生物学的可识别影响的研究结果。
本论文首次应用代谢组学方法研究精油抗焦虑功效。利用色谱-质谱联用技术,包括气相色谱／飞行时间质谱(GC-TOF/MS)和超高效液相色谱-质谱(UPLC/Q-TOF-MS)技术,对大鼠尿液和脑组织样本以及人的尿液样本进行了代谢物的高通量检测,并完成了数据的统计学分析。
主要内容和结果如下：
1、采用大鼠非条件反射焦虑模型——高架十字迷宫(EPM)对精油香气对于动物的焦虑行为的影响进行评价,利用色谱质谱联用技术测试尿液和脑组织样本,分析高架十字迷宫(EPM)组与闻香-EPM组的代谢谱差异。研究捕捉到了基于大鼠高架十字迷宫焦虑模型(EPM)的精油抗焦虑作用以及香气诱发的生物体内代谢变化。与未嗅闻香气的EPM大鼠相比,嗅闻精油香气的EPM大鼠代谢物的变化减弱,表明闻香-EPM组的代谢水平波动程度较小,说明精油香气的吸入缓解了大鼠因EPM实验所产生的代谢波动。精油香气对于代谢变化的平抑作用与EPM行为学评价中闻香-EPM组的开放臂进入时间比例OT%和开放臂进入次数比例OE%上升的结果是一致的。
2、通过志愿者精油闻香实验,采收闻香期前后的尿液样本,利用GC-TOF/MS及UPLC/Q-TOF-MS分析测试平台技术,结合SCL-90状态自测量表,对人体在精油香气的作用下的情绪变化和体内代谢物差异进行分析。结果表明,利用非监督的PCA方法,较难分离闻香期前后志愿者尿液中的代谢物谱图,运用PLS-DA方法进行分析,可以明显地观察到闻香前后代谢谱存在的差异。进一步运用GC-TOF/MS和UPLC/Q-TOF-MS数据进行联合分析,发现了不同的人对于精油香气扰动的代谢物表现差异是非常大的,可以明显地呈现出精油香气易感和不易感两个群体。对发现的29个重要差异性代谢物进行分析,表明焦虑情绪的变化可能与能量代谢、氨基酸代谢以及有机酸代谢、肠道菌群代谢等代谢通路的变化具有相关性。
3、利用气相色谱(GC)和气相色谱-质谱联用(GC-MS),结合保留指数的计算和匹配,对精油的成分进行分析。定性定量出34种化合物,相对含量为92.81%,其中主要成分为醇类34.14%、萜烯类27.77%、酯类26.27%。
本论文创新性地基于代谢组学对精油抗焦虑功效进行了研究,本研究的意义在于首次表明通过代谢组学技术可以捕捉精油香气对于生物体所产生的微妙的代谢变化,为探寻焦虑相关行为的作用通路和进一步研究精油的抗焦虑机理奠定了基础和指引了方向。
Anxiety disorders are the most prevalent psychiatric disorders and affect a greatnumber of people worldwide. Essential oils, take effects through inhalation or topicalapplication, are believed to enhance physical, emotional, and spiritual well-being.Although clinical studies suggest that the use of essential oils may have therapeuticpotential, evidence for the efficacy of essential oils in treating medical conditionsremains poor, with a particular lack of studies employing rigorous analytical methodsthat capture its identifiable impact on human biology.
In this dissertation, it is the first time to apply metabonomics method to study theanxiolytic effect of essential oils and reveal the aromas-induced metabolic changes.We examined the metabolites of different bio-samples, including samples of humenand Wistar rats using GC-TOF/MS&UPLC/Q-TOF-MS, and carried out statisticalanalysis of the data.
Main methods and results:
1. It reveals the aromas-induced metabolic changes and the anxiolytic effect ofaromas in elevated plus maze (EPM) induced anxiety model rats. The significantalteration of metabolites in the EPM group was attenuated by aromas treatment,concurrent with the behavioral improvement with significantly increased openarms time and open arms entries.
2. We examined the metabolites of urine samples of humen before and after10-daysmelling aroma of essential oils, combined with the result of Symptom Checklist90. The results show that using the unsupervised PCA method can hardly get aseparation trend. It can be clearly observed in the metabolic profile of thedifferences between the two group using PLS-DA method. From the GC&UPLCcombined metabolomics results, it was found some people’s metabolic profileswere not changed much after aromatherapy while others were significantly altered.There are29important differences in metabolite relating to anxiety and the metabolic path changes including energy metabolism, amino acid metabolism,organic acid metabolism and glu flora metabolism.
3. Using gas chromatography (GC) and gas chromatography mass spectrometry(GC-MS) combined with Temperature-programmed retention indices onessential oil composition analysis. Qualitative and quantitative analysis of34kinds of compounds, the relative content of92.81%, which is the main componentof alcohols, esters, terpenes34.14%27.77%26.27%respectively.
This study demonstrates, for the first time, that the metabonomics approach cancapture the subtle metabolic changes resulting from exposure to essential oils andprovide the basis for pinpointing affected pathways in anxiety-related behavior, whichwill lead to an improved mechanistic understanding of anxiolytic effect of essentialoils.
本论文首次应用代谢组学方法研究精油抗焦虑功效。利用色谱-质谱联用技术,包括气相色谱／飞行时间质谱(GC-TOF/MS)和超高效液相色谱-质谱(UPLC/Q-TOF-MS)技术,对大鼠尿液和脑组织样本以及人的尿液样本进行了代谢物的高通量检测,并完成了数据的统计学分析。
主要内容和结果如下：
1、采用大鼠非条件反射焦虑模型——高架十字迷宫(EPM)对精油香气对于动物的焦虑行为的影响进行评价,利用色谱质谱联用技术测试尿液和脑组织样本,分析高架十字迷宫(EPM)组与闻香-EPM组的代谢谱差异。研究捕捉到了基于大鼠高架十字迷宫焦虑模型(EPM)的精油抗焦虑作用以及香气诱发的生物体内代谢变化。与未嗅闻香气的EPM大鼠相比,嗅闻精油香气的EPM大鼠代谢物的变化减弱,表明闻香-EPM组的代谢水平波动程度较小,说明精油香气的吸入缓解了大鼠因EPM实验所产生的代谢波动。精油香气对于代谢变化的平抑作用与EPM行为学评价中闻香-EPM组的开放臂进入时间比例OT%和开放臂进入次数比例OE%上升的结果是一致的。
2、通过志愿者精油闻香实验,采收闻香期前后的尿液样本,利用GC-TOF/MS及UPLC/Q-TOF-MS分析测试平台技术,结合SCL-90状态自测量表,对人体在精油香气的作用下的情绪变化和体内代谢物差异进行分析。结果表明,利用非监督的PCA方法,较难分离闻香期前后志愿者尿液中的代谢物谱图,运用PLS-DA方法进行分析,可以明显地观察到闻香前后代谢谱存在的差异。进一步运用GC-TOF/MS和UPLC/Q-TOF-MS数据进行联合分析,发现了不同的人对于精油香气扰动的代谢物表现差异是非常大的,可以明显地呈现出精油香气易感和不易感两个群体。对发现的29个重要差异性代谢物进行分析,表明焦虑情绪的变化可能与能量代谢、氨基酸代谢以及有机酸代谢、肠道菌群代谢等代谢通路的变化具有相关性。
3、利用气相色谱(GC)和气相色谱-质谱联用(GC-MS),结合保留指数的计算和匹配,对精油的成分进行分析。定性定量出34种化合物,相对含量为92.81%,其中主要成分为醇类34.14%、萜烯类27.77%、酯类26.27%。
本论文创新性地基于代谢组学对精油抗焦虑功效进行了研究,本研究的意义在于首次表明通过代谢组学技术可以捕捉精油香气对于生物体所产生的微妙的代谢变化,为探寻焦虑相关行为的作用通路和进一步研究精油的抗焦虑机理奠定了基础和指引了方向。
Anxiety disorders are the most prevalent psychiatric disorders and affect a greatnumber of people worldwide. Essential oils, take effects through inhalation or topicalapplication, are believed to enhance physical, emotional, and spiritual well-being.Although clinical studies suggest that the use of essential oils may have therapeuticpotential, evidence for the efficacy of essential oils in treating medical conditionsremains poor, with a particular lack of studies employing rigorous analytical methodsthat capture its identifiable impact on human biology.
In this dissertation, it is the first time to apply metabonomics method to study theanxiolytic effect of essential oils and reveal the aromas-induced metabolic changes.We examined the metabolites of different bio-samples, including samples of humenand Wistar rats using GC-TOF/MS&UPLC/Q-TOF-MS, and carried out statisticalanalysis of the data.
Main methods and results:
1. It reveals the aromas-induced metabolic changes and the anxiolytic effect ofaromas in elevated plus maze (EPM) induced anxiety model rats. The significantalteration of metabolites in the EPM group was attenuated by aromas treatment,concurrent with the behavioral improvement with significantly increased openarms time and open arms entries.
2. We examined the metabolites of urine samples of humen before and after10-daysmelling aroma of essential oils, combined with the result of Symptom Checklist90. The results show that using the unsupervised PCA method can hardly get aseparation trend. It can be clearly observed in the metabolic profile of thedifferences between the two group using PLS-DA method. From the GC&UPLCcombined metabolomics results, it was found some people’s metabolic profileswere not changed much after aromatherapy while others were significantly altered.There are29important differences in metabolite relating to anxiety and the metabolic path changes including energy metabolism, amino acid metabolism,organic acid metabolism and glu flora metabolism.
3. Using gas chromatography (GC) and gas chromatography mass spectrometry(GC-MS) combined with Temperature-programmed retention indices onessential oil composition analysis. Qualitative and quantitative analysis of34kinds of compounds, the relative content of92.81%, which is the main componentof alcohols, esters, terpenes34.14%27.77%26.27%respectively.
This study demonstrates, for the first time, that the metabonomics approach cancapture the subtle metabolic changes resulting from exposure to essential oils andprovide the basis for pinpointing affected pathways in anxiety-related behavior, whichwill lead to an improved mechanistic understanding of anxiolytic effect of essentialoils.
引文
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