| 引用本文:姬党通1，2，陈玉莹1，2，李欠1，2.荧光成像技术用于独活根中香豆素类成分积累规律研究[J].世界中医药,2022,(24):. |
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| 荧光成像技术用于独活根中香豆素类成分积累规律研究 |
| Accumulation Pattern of Coumarins in Angelica pubescens Root Based on Fluorescence Imaging |
| 投稿时间：2022-01-24 |
| DOI：10.3969/j.issn.1673-7202.2022.24.001 |
| 中文关键词: 独活 香豆素 冰冻切片 组织化学定位 荧光成像技术 分泌腔 |
| English Keywords:Angelica pubescens root Coumarins Frozen section Histochemical localization Fluorescence imaging Secretory cavity |
| 基金项目:国家自然科学基金项目（31860102）；甘肃农业大学人才引进专项（GSAU-RCZX201704）；甘肃省优秀研究生“创新之星”项目（2021CXZX-399） |
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| 中文摘要: |
| 目的：建立独活冰冻切片的新方法，并利用荧光成像技术对不同时期独活根中的香豆素类成分进行组织化学定位、定量研究，明确独活中活性成分香豆素的组织分布及其动态积累规律。方法：以不同时期的独活根为材料，考察不同保护剂、不同切片厚度对独活根最佳制片的影响，以此建立独活根的最佳制片条件。此外，利用显微成像测量技术对不同部位分泌组织进行统计分析，并结合荧光成像对香豆素成分集中分布部位的荧光强度进行测算，以荧光强度对不同时期的独活根中香豆素成分进行相对定量研究，分析不同时期香豆素成分含量的动态变化规律。结果：独活根经15%甘油保护剂的处理，切片厚度为25 μm时切片效果最佳。通过组织化学定位显示，独活香豆素类成分主要分布在韧皮部的分泌腔及分泌道中。不同时期香豆素类成分总体变化趋势呈S形增长，8月、10月含量变化明显，10月香豆素含量达到最大。结论：首次使用了独活根冰冻切片及荧光成像新方法，研究了香豆素成分的产生、分布以及积累规律。发现独活香豆素成分主要分布在韧皮部的分泌腔中；不同时期独活根中香豆素成分动态积累变化趋势呈S形增长，且10月香豆素含量积累达到最大，为拓宽中药分析提供了新思路。 |
| English Summary: |
| To establish a new method for frozen section of Angelicae pubescens root，and study the histochemical localization and conduct quantitative analysis of coumarins in A.pubescens root at different growth stages by fluorescence imaging，to clarify the tissue distribution and dynamic accumulation pattern of active component coumarins in A.pubescens root.Methods:With A.pubescens root at different growth stage used as the research material，the effects of different protective agents and section thickness on the optimal section of A.pubescens root were investigated，and the optimum conditions for sections were established.In addition，microscopic imaging technology was used for statistical analysis of secreted tissues in different parts，and fluorescence imaging was combined to measure the fluorescence intensity of the parts where coumarins were concentrated.The relative quantitative analysis of coumarins in A.pubescens root at different growth stages was carried out by fluorescence intensity，and their dynamic change pattern was analyzed.Results:The optimal section effect of A.pubescens root was obtained when the section thickness was 25 μm and under the treatment with 15% glycerol protectant.Histochemical localization showed that coumarins were mainly distributed in the secretory cavity and secretory tract of the phloem.The overall change trend of coumarins in A.pubescens root at different growth stages presented a S-shaped increase，with obvious content change in August and October and the maximum content in October.Conclusion:The frozen sectioning and fluorescence imaging of A.pubescens root were established for the first time，and the generation，distribution and accumulation of coumarins were studied.It was found that coumarins were mainly distributed in the secretory cavity of the phloem.The dynamic change trend of coumarins in A.pubescens root at different growth stages showed a S-shaped increase，and the accumulation of coumarins reached the maximum in October.This paper provides a new idea for expanding the analysis of traditional Chinese medicine. |
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