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金莲花不同溶剂提取物抗心肌细胞缺氧/复氧损伤的谱-效关系

研究 Δ

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南敏伦 ，孙 琦 ，杨 振 ，司学玲 ，马春霞 ，白 雪 ，赫玉芳 （1.吉林省中医药科学院植物化学研究所，长
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春 130012；2.吉林正容医药发展有限责任公司，吉林 敦化 133700；3.修正药业集团长春高新制药有限公司，
长春 130012；4.吉林农业大学中药材学院，长春 130118；5.长春中医药大学健康管理学院，长春 130117）
中图分类号 R285.5 文献标志码 A 文章编号 1001-0408（2020）18-2219-05
DOI 10.6039/j.issn.1001-0408.2020.18.08
摘 要 目的：研究金莲花11种不同溶剂提取物抗心肌细胞缺氧/复氧损伤的谱-效关系。方法：利用高效液相色谱-串联质谱法
建立金莲花11种不同溶剂提取物的指纹图谱，通过与对照品、文献信息比对，指认共有峰对应的化合物。采用MTT法检测金莲花
11种不同溶剂提取物对缺氧/复氧损伤大鼠心肌细胞H9c2存活率的影响，采用酶联免疫吸附测定法检测细胞中丙二醛（MDA）含
量、活性氧（ROS）水平和上清液中乳酸脱氢酶（LDH）含量。通过灰色关联度分析法（GRA）和偏最小二乘法（PLS）联合分析共有
峰对应成分与抗心肌细胞缺氧/复氧损伤（药效）的谱-效关系。结果：金莲花11种不同溶剂提取物的指纹图谱中有22个共有峰，指
认出22个化合物。与缺氧/复氧损伤组比较，缺氧/复氧+S1～S6、S9、S10组细胞的存活率均显著升高，MDA含量、ROS水平、LDH
含量均显著降低（P＜0.05）；缺氧/复氧+S8组细胞的ROS水平、LDH含量均显著降低（P＜0.05）。22个化合物与药效的GRA关联
度均大于0.8；除峰1、2、7、13、14、21外，其余峰对应化合物与药效的PLS分析回归系数均大于0（即成正相关）；贡献排名前9位的
共有峰均为峰 6＞11＞4＞5＞8＞9＞12＞10＞15。结论：荭草苷（峰 6）、牡荆苷（峰 11）、荭草素-2″-O-β-L-半乳糖苷（峰 4）、荭草
素-2″-O-β-D-吡喃木糖苷（峰 5）、槲皮素-3-O-吡喃葡萄糖苷（峰 8）、牡荆素-2″-O-β-L-半乳糖苷（峰 9）、金丝桃苷（峰 12）、牡荆
素-2″-O-β-D-吡喃木糖苷（峰10）、2″-O-（2″′ -甲基丁酰基）-荭草苷（峰15）可能是金莲花抗心肌细胞缺氧/复氧损伤的主要成分。
关键词 金莲花；心肌细胞H9c2；缺氧/复氧损伤；谱-效关系；灰色关联度分析法；偏最小二乘法

Study on the Spectrum-effect Relationship of Different Solvent Extracts from Trollius chinensis against
Hypoxia/reoxygenation Injury of Cardiomyocyte
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NAN Minlun ，SUN Qi ，YANG Zhen ，SI Xueling ，MA Chunxia ，BAI Xue ，HE Yufang （1. Institute of
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Phytochemistry，Jilin Academy of Chinese Medicine Sciences，Changchun 130012，China；2. Jilin Zhengrong
Medical Development Co.，Ltd.，Jilin Dunhua 133700，China；3. Xiuzheng Pharmaceutical Group Changchun
High-tech Pharmaceutical Co.， Ltd.， Changchun 130012， China；4. College of TCM， Jilin Agricultural
University，Changchun 130118，China；5. College of Health Management，Changchun University of TCM，
Changchun 130117，China）
ABSTRACT OBJECTIVE：To study spectrum-effect relationship of 11 different solvent extracts from Trollius chinensis against
hypoxia/reoxygenation injury of cardiomyocyte. METHODS：HPLC-MS/MS method was used to establish the fingerprints of 11
different solvent extracts from T. chinensis，the compounds corresponding to the common peaks were identified by comparing with
the substance control and literature information. MTT assay was used to detect the effects of 11 different solvent extracts from T.
chinensis on the survival rate of rat myocardial H9c2 cells injured by hypoxia/reoxygenation. The MDA content，ROS level in cells
and LDH content in the supernatant were detected by ELISA. GRA and PLS method were used to analyze the spectrum-effect
relationship between the compounds corresponding to common peak and anti-hypoxia/reoxygenation injury of cardiomyocyte（drug
effect）. RESULTS：There were 22 common peaks in 11 different solvent extracts from T. chinensis，and 22 compounds were
identified. Compared with hypoxia/reoxygenation injury group，survival rate of hypoxia/reoxygenation injury+S1-S6，S9 and S10
groups were increased significantly，while MDA content，ROS level and LDH content were decreased significantly（P＜0.05）；
ROS level and LDH content of hypoxia/reoxygenation injury + S8 group were decreased significantly（P＜0.05）. The r of GRA
analysis of 22 compounds with drug effects were all higher
Δ 基金项目：吉林省科技发展计划项目（No.20150203010YY）；吉
than 0.8. Except for peaks 1，2，7，13，14 and 21，r of PLS
林省中医药科技项目（No.2019031）
analysis of rest peaks with drug effects were higher than 0
＊副研究员，硕士。研究方向：中药化学成分提取、分离及产品开
（being positive correlation）. Top 9 common peaks in the list
发。电话：0431-86058683。E-mial：nml2000@163.com
# 通信作者：教授，博士。研究方向：中药化学成分提取、分离及 of contribution rate were peak 6＞11＞4＞5＞8＞9＞12＞10＞
产品开发。电话：0431-86058683。E-mial：hyf_1992@163.com 15. CONCLUSIONS：Orientin（peak 6），vitexin（peak 11），

中国药房 2020年第31卷第18期 China Pharmacy 2020 Vol. 31 No. 18 ·2219 ·