中国药物警戒 ›› 2023, Vol. 20 ›› Issue (6): 623-628.
DOI: 10.19803/j.1672-8629.20230110

• 何首乌质量安全及药效研究专栏 • 上一篇    下一篇

红参-制何首乌药对对神经细胞保护作用的谱效关系

刘晶晶1, 戴忠1△, 杨建波1, 王莹1, 徐蓓蕾2, 刘越3, 汪祺1#, 马双成1,*   

  1. 1中国食品药品检定研究院中药民族药检定所,北京 100050;
    2哈尔滨商业大学药学院,黑龙江 哈尔滨 150000;
    3北京中医药大学药学院,北京 100871
  • 收稿日期:2023-02-27 出版日期:2023-06-15 发布日期:2023-06-15
  • 通讯作者: * 马双成,男,研究员·博导,中药民族药物质基础与质量安全评价研究。E-mail：msc@nifdc.org.cn。#为共同通信作者。
  • 作者简介:刘晶晶,女,硕士,助理研究员,中药质量控制研究。△为共同第一作者。
  • 基金资助:
    国家自然科学基金资助项目（81973476）

Spectrum-fect relationship of Ginseng Radix et Rhizoma Rubra and Polygonum Multiflorum Radix Preparata medicinal pair in protection of nerve cells

LIU Jingjing1, DAI Zhong1△, YANG Jianbo1, WANG Ying1, XU Beilei2, LIU Yue3, WANG Qi1#, MA Shuangcheng1,*   

  1. 1National Institutes for Food and Drug Control, Institute for Control of Chinese Traditional Medicine and Ethnic Medicine, Beijing 100050, China;
    2School of Pharmacy, Harbin University of Commerce, Harbin Heilongjiang 150000, China;
    3School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100871, China
  • Received:2023-02-27 Online:2023-06-15 Published:2023-06-15

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摘要： 目的 建立红参-制何首乌药对不同提取工艺的超高效液相色谱（UHPLC）指纹图谱,并研究其对Aβ1-42诱导HT22细胞模型保护作用的谱效关系。方法 制备红参-制何首乌药对30%乙醇回流提取物、30%乙醇渗漉提取物、70%乙醇回流提取物、70%乙醇渗漉提取物和水提取物。采用UHPLC法进行测定并结合《中药色谱指纹图谱相似度评价系统》（2012A版）建立不同工艺提取物的UHPLC指纹图谱,并进行共有峰指认。采用Aβ1-42诱导海马神经元HT22细胞,利用MTT法评估不同工艺提取物对模型的保护,考察红参-制何首乌药对不同提取工艺对神经细胞的保护作用;采用灰色关联度分析法、偏最小二乘法（PLS）分析不同提取工艺UHPLC指纹图谱中共有峰与神经细胞保护的相关性。结果 红参-制何首乌药对不同提取工艺有19个共有峰,通过化学对照品指认13个成分。经灰色关联度分析发现,14个共有峰与神经细胞保护的关联度均大于0.6,呈相关性。经PLS分析发现10个成分对神经细胞保护作用的影响较大。结论 建立红参-制何首乌药对不同提取工艺UHPLC指纹图谱并指认了19个共有峰成分;大黄素-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、大黄素、大黄素甲醚、20(S)-人参皂苷 Rg2、人参皂苷Ro、人参皂苷Rd和14、17、18号峰所代表的化学成分可能是神经细胞保护作用的药效物质。

关键词: 红参-制何首乌药对, 神经细胞保护, 指纹图谱, 谱效关系, Aβ1-42, HT-22 细胞, 灰色关联度, 偏最小二乘法

Abstract: Objective To establish ultrahigh performance liquid chromatography (UHPLC) fingerprints of Ginseng Radix et Rhizoma Rubra(GR) and Polygonum Multiflorum Radix Preparata (PMRP) for different extraction processes, and to study the spectrum-effect relationship with the protective effect against Aβ1-42 induced HT-22 cell injury. Methods 30% ethanol reflux extracts, 30% ethanol percolation extracts, 70% ethanol reflux extracts, 70% ethanol percolation extracts and water extracts from GR and PMRP were prepared. UHPLC fingerprints for different extraction processes were established using the UHPLC method combined with the Similarity Evaluation System of TCM Chromatogramtic Fingerprint (2012A) before the common peaks were identified. Aβ1-42 was used to induce HT22 cells of hippocampal neurons. The protective effect of extracts using different processes on the model was evaluated with the MTT method. Using the protective effect against Aβ1-42 induced HT-22 cell injury as indexes, the properties of extracts using different processes were investigated. The correlations between the common peaks of UHPLC fingerprints and the indexes of analgesic and protective effect were analyzed via grey correlation analysis and the partial least square（PLS）method. Results There were 19 common peaks for different extraction processes of GR and PMRP, and 13 components were identified by reference comparison. Grey correlation analysis found that the degree of correlation between the 14 common peaks and neural cell protection exceeded 0.6, suggesting good correlations. PLS analysis indicated that peaks 6, 7, 8, 9, 10, 12, 13, 14, 17 and 18 had a greater impact on the protection of nerve cells. Conclusion UHPLC fingerprints for different extraction processes of GR and PMRP are established and 19 common peak components are identified. Emodin-8-O-β-D-glucoside, emodin methyl ether-8-O-β-D-glucoside, emodin, emodin methyl ether, 20(S)-ginsenoside Rg2, ginsenoside Ro, ginsenoside Rd and peaks 14, 17 and 18 may be the effective substances of neuroprotection.

Key words: Ginseng Radix et Rhizoma Rubra and Polygonum Multiflorum Radix Preparata medicinal pair, neuroprotection, fingerprint, spectrum-effect relationship, Aβ1-42, HT-22 cells, grey correlation analysis, PLS

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