Article(id=1198628604004365308, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198628599835227075, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2022-1340, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1670342400000, receivedDateStr=2022-12-07, revisedDate=1672329600000, revisedDateStr=2022-12-30, acceptedDate=null, acceptedDateStr=null, onlineDate=1763704928637, onlineDateStr=2025-11-21, pubDate=1686499200000, pubDateStr=2023-06-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1763704928637, onlineIssueDateStr=2025-11-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1763704928637, creator=13701087609, updateTime=1763704928637, updator=13701087609, issue=Issue{id=1198628599835227075, tenantId=1146029695717560320, journalId=1189982191388893191, year='2023', volume='58', issue='6', pageStart='0', pageEnd='1724', issueExtLink='null', onlineDate='null', pubDate='1686499200000', pubDateStr='2023-06-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1763704927643, creator='13701087609', updateTime=1766137690373, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1208832346359468483, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198628599835227075, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1208832346359468484, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198628599835227075, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1629, endPage=1633, ext={EN=ArticleExt(id=1198628604390240272, articleId=1198628604004365308, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Flavonoid glycosides from the water extract of Artemisia annua L., columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

Twenty one flavonoid glycosides were isolated and purified from n-butanol portion of the water extract of A. annua by various chromatographic techniques such as HP-20 macroporous adsorption resin, silica gel, ODS, Sephadex LH-20 gel column chromatography and preparative high performance liquid chromatography. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as axillarin-7-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside (1), orientin (2), apigenin-6-C-β-D-glucopyranosyl-8-C-β-L-arabinopyranoside (3), apigenin-6-C-β-D-galactopyranosyl-8-C-β-L-arabinopyranoside (4), apigenin-6-C-β-L-arabinopyranosyl-8-C-β-D-glucopyranoside (5), apigenin-6-C-α-L-arabinofuranosyl-8-C-β-D-glucopyranoside (6), quercetin-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (7), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (8), vicenin-2 (9), patuletin-7-O-β-D-glucopyranoside (10), luteolin-6-C-glucopyranoside (11), vitexin (12), kaempferol-3-O-β-galactopyranosyl-(1→2)-β-glucopyranoside (13), quercetin-7-O-β-D-glucopyranoside (14), patuletin-3-O-β-D-glucopyranoside (15), 7-O-methyl-quercetagetin-6-O-β-D-glucopyranoside (16), quercetin-3-O-β-D-glucopyranoside (17), nepitrin (18), rutin (19), kaempferol-3-O-β-sophoroside (20), and patuletin-3-O-rutinoside (21). Compound 1 is a new compound, compounds 2, 4, 6, 7, 10, 11, 13, 15, 16, 18, 20 and 21 are isolated from A. annua for the first time. In the anti-inflammatory assay, compound 1 inhibited the release of IL-6 from LPS-induced RAW264.7 cells to significantly degrees with the high (100 μmol·L-1), medium (50 μmol·L-1), low (25 μmol·L-1) concentration.

, authors=null, authorsList=Qi-guo WU, Ming-hui FAN, Le-yi HUANG, Yong-li WANG, Gui-xin CHOU, authorCompany=null, correspAuthors=Gui-xin CHOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2023 Acta Pharmaceutica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1198628605392678994, articleId=1198628604004365308, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=黄花蒿水提物黄酮苷类成分研究, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

采用HP-20大孔吸附树脂、MCI柱色谱、硅胶柱色谱、中压液相色谱、葡聚糖凝胶Sephadex LH-20柱色谱及制备液相色谱等多种色谱技术, 从黄花蒿水提物的正丁醇部位分离得到21个黄酮苷类化合物。根据化合物的理化性质和波谱数据确定了它们的结构, 分别为甲氧基万寿菊素-7-O-β-D-吡喃木糖-(1→6)-β-D-吡喃葡萄糖苷(1)、荭草苷(2)、芹菜素-6-C-β-D-吡喃葡萄糖-8-C-β-L-吡喃阿拉伯糖苷(3)、芹菜素-6-C-β-D-吡喃半乳糖-8-C-β-L-吡喃阿拉伯糖苷(4)、芹菜素-6-C-β-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(5)、芹菜素-6-C-α-L-呋喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(6)、槲皮素-3-O-β-D-吡喃葡萄糖苷-(1→2)-β-D-吡喃葡萄糖苷(7)、芹菜素-6-C-α-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(8)、芹菜素-6, 8-二-C-吡喃葡萄糖苷(9)、万寿菊素-7-O-β-D-吡喃葡萄糖苷(10)、木犀草素-6-C-吡喃葡萄糖苷(11)、牡荆素(12)、山柰酚-3-O-β-吡喃半乳糖-(1→2)-β-吡喃葡萄糖苷(13)、槲皮素-7-O-β-D-吡喃葡萄糖苷(14)、万寿菊素-3-O-β-D-吡喃葡萄糖苷(15)、7-甲氧基-栎草亭-6-O-β-D-吡喃葡萄糖苷(16)、槲皮素-3-O-β-D-吡喃葡萄糖苷(17)、假荆芥属苷(18)、芦丁(19)、山柰酚-3-O-β-槐糖苷(20) 和万寿菊素-3-O-β-D-芦丁糖苷(21)。其中, 化合物1为新化合物, 化合物24671011131516182021为首次从黄花蒿中分离得到。通过ELISA法检测新化合物对LPS诱导的RAW264.7细胞IL-6蛋白表达实验测定了新化合物的抗炎活性, 结果表明化合物1在高(100 μmol·L-1)、中(50 μmol·L-1)、低(25 μmol·L-1) 浓度均能表现出显著抑制IL-6蛋白的生成与表达。

, authors=null, authorsList=吴其国, 范明惠, 黄乐怡, 王永丽, 侴桂新, authorCompany=null, correspAuthors=侴桂新, authorNote=null, correspAuthorsNote=
*侴桂新, Tel: 86-21-51322623, E-mail:
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PositionδH (J in Hz)δC
2158.7
3139.5
4180.4
5153.8
6133.9
7157.8
86.92, s95.5
9153.4
10108.2
1′122.8
2′7.70, d (2.2)116.8
3′146.4
4′150.1
5′6.95, d (8.5)116.5
6′7.58, dd (8.5, 2.2)122.7
1″5.12, d (7.3)102.0
2″3.56-3.61, m74.7
3″3.49-3.56, m77.7
4″3.45-3.52, m71.1
5″3.79-3.72, m77.3
6″a4.18, dd (11.3, 1.7)69.9
6″b3.80-3.86, m
1‴4.32, d (7.0)105.4
2‴3.23-3.27, m74.8
3‴3.27-3.32, m77.7
4‴3.45-3.52, m71.0
5‴a3.89-3.83, m66.9
5‴b3.16, dd (11.4, 10.0)
3-OCH33.82, s60.5
6-OCH33.91, s61.5
), ArticleFig(id=1199640570592325900, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198628604004365308, language=CN, label=Table 1, caption=

The 1H NMR (400 MHz) and 13C NMR (100 MHz) data of compound 1 (measured in methanol-d4)

, figureFileSmall=null, figureFileBig=null, tableContent=
PositionδH (J in Hz)δC
2158.7
3139.5
4180.4
5153.8
6133.9
7157.8
86.92, s95.5
9153.4
10108.2
1′122.8
2′7.70, d (2.2)116.8
3′146.4
4′150.1
5′6.95, d (8.5)116.5
6′7.58, dd (8.5, 2.2)122.7
1″5.12, d (7.3)102.0
2″3.56-3.61, m74.7
3″3.49-3.56, m77.7
4″3.45-3.52, m71.1
5″3.79-3.72, m77.3
6″a4.18, dd (11.3, 1.7)69.9
6″b3.80-3.86, m
1‴4.32, d (7.0)105.4
2‴3.23-3.27, m74.8
3‴3.27-3.32, m77.7
4‴3.45-3.52, m71.0
5‴a3.89-3.83, m66.9
5‴b3.16, dd (11.4, 10.0)
3-OCH33.82, s60.5
6-OCH33.91, s61.5
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黄花蒿水提物黄酮苷类成分研究
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吴其国 1, 2, 3 , 范明惠 1, 2 , 黄乐怡 1, 2 , 王永丽 1, 2 , 侴桂新 1, 2, *
药学学报 | 研究论文 2023,58(6): 1629-1633
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药学学报 |研究论文 2023 , 58 (6) : 1629 -1633
黄花蒿水提物黄酮苷类成分研究
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吴其国1, 2, 3, 范明惠1, 2, 黄乐怡1, 2, 王永丽1, 2, 侴桂新1, 2, *
作者信息
  • 1.上海中医药大学中药研究所, 上海 201203
  • 2.上海中药标准化研究中心, 上海 201203
  • 3.安庆医药高等专科学校药学院, 安徽 安庆 246052
通讯作者:
*侴桂新, Tel: 86-21-51322623, E-mail:
Flavonoid glycosides from the water extract of Artemisia annua L.
Qi-guo WU1, 2, 3, Ming-hui FAN1, 2, Le-yi HUANG1, 2, Yong-li WANG1, 2, Gui-xin CHOU1, 2, *
Affiliations
  • 1. Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • 2. Shanghai R & D Centre for Standardization of Chinese Medicines, Shanghai 201203, China
  • 3. Department of Pharmacy, Anqing Medical College, Anqing 246052, China
出版时间: 2023-06-12 doi: 10.16438/j.0513-4870.2022-1340
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采用HP-20大孔吸附树脂、MCI柱色谱、硅胶柱色谱、中压液相色谱、葡聚糖凝胶Sephadex LH-20柱色谱及制备液相色谱等多种色谱技术, 从黄花蒿水提物的正丁醇部位分离得到21个黄酮苷类化合物。根据化合物的理化性质和波谱数据确定了它们的结构, 分别为甲氧基万寿菊素-7-O-β-D-吡喃木糖-(1→6)-β-D-吡喃葡萄糖苷(1)、荭草苷(2)、芹菜素-6-C-β-D-吡喃葡萄糖-8-C-β-L-吡喃阿拉伯糖苷(3)、芹菜素-6-C-β-D-吡喃半乳糖-8-C-β-L-吡喃阿拉伯糖苷(4)、芹菜素-6-C-β-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(5)、芹菜素-6-C-α-L-呋喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(6)、槲皮素-3-O-β-D-吡喃葡萄糖苷-(1→2)-β-D-吡喃葡萄糖苷(7)、芹菜素-6-C-α-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(8)、芹菜素-6, 8-二-C-吡喃葡萄糖苷(9)、万寿菊素-7-O-β-D-吡喃葡萄糖苷(10)、木犀草素-6-C-吡喃葡萄糖苷(11)、牡荆素(12)、山柰酚-3-O-β-吡喃半乳糖-(1→2)-β-吡喃葡萄糖苷(13)、槲皮素-7-O-β-D-吡喃葡萄糖苷(14)、万寿菊素-3-O-β-D-吡喃葡萄糖苷(15)、7-甲氧基-栎草亭-6-O-β-D-吡喃葡萄糖苷(16)、槲皮素-3-O-β-D-吡喃葡萄糖苷(17)、假荆芥属苷(18)、芦丁(19)、山柰酚-3-O-β-槐糖苷(20) 和万寿菊素-3-O-β-D-芦丁糖苷(21)。其中, 化合物1为新化合物, 化合物24671011131516182021为首次从黄花蒿中分离得到。通过ELISA法检测新化合物对LPS诱导的RAW264.7细胞IL-6蛋白表达实验测定了新化合物的抗炎活性, 结果表明化合物1在高(100 μmol·L-1)、中(50 μmol·L-1)、低(25 μmol·L-1) 浓度均能表现出显著抑制IL-6蛋白的生成与表达。

黄花蒿  /  水提物  /  黄酮  /  抗炎

Twenty one flavonoid glycosides were isolated and purified from n-butanol portion of the water extract of A. annua by various chromatographic techniques such as HP-20 macroporous adsorption resin, silica gel, ODS, Sephadex LH-20 gel column chromatography and preparative high performance liquid chromatography. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as axillarin-7-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside (1), orientin (2), apigenin-6-C-β-D-glucopyranosyl-8-C-β-L-arabinopyranoside (3), apigenin-6-C-β-D-galactopyranosyl-8-C-β-L-arabinopyranoside (4), apigenin-6-C-β-L-arabinopyranosyl-8-C-β-D-glucopyranoside (5), apigenin-6-C-α-L-arabinofuranosyl-8-C-β-D-glucopyranoside (6), quercetin-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (7), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (8), vicenin-2 (9), patuletin-7-O-β-D-glucopyranoside (10), luteolin-6-C-glucopyranoside (11), vitexin (12), kaempferol-3-O-β-galactopyranosyl-(1→2)-β-glucopyranoside (13), quercetin-7-O-β-D-glucopyranoside (14), patuletin-3-O-β-D-glucopyranoside (15), 7-O-methyl-quercetagetin-6-O-β-D-glucopyranoside (16), quercetin-3-O-β-D-glucopyranoside (17), nepitrin (18), rutin (19), kaempferol-3-O-β-sophoroside (20), and patuletin-3-O-rutinoside (21). Compound 1 is a new compound, compounds 2, 4, 6, 7, 10, 11, 13, 15, 16, 18, 20 and 21 are isolated from A. annua for the first time. In the anti-inflammatory assay, compound 1 inhibited the release of IL-6 from LPS-induced RAW264.7 cells to significantly degrees with the high (100 μmol·L-1), medium (50 μmol·L-1), low (25 μmol·L-1) concentration.

Artemisia annua L.  /  water extract  /  flavonoid  /  anti-inflammatory
吴其国, 范明惠, 黄乐怡, 王永丽, 侴桂新. 黄花蒿水提物黄酮苷类成分研究. 药学学报, 2023 , 58 (6) : 1629 -1633 . DOI: 10.16438/j.0513-4870.2022-1340
Qi-guo WU, Ming-hui FAN, Le-yi HUANG, Yong-li WANG, Gui-xin CHOU. Flavonoid glycosides from the water extract of Artemisia annua L.[J]. Acta Pharmaceutica Sinica, 2023 , 58 (6) : 1629 -1633 . DOI: 10.16438/j.0513-4870.2022-1340
崇明苦草是上海市崇明地区世代相传的一种药草, 因其疗效显著而家喻户晓。其基原植物为菊科植物黄花蒿Artemisia annua L., 具有清热解毒、消肿止痛的功效, 并有多种食疗作用[1]。虽然与中药青蒿为同一基原, 但有别于青蒿的是, 崇明苦草一般于每年3月份播种, 10~11月份果实成熟后采收, 取其带成熟果实的茎枝, 晾干, 储存3年, 作为“崇明苦草”应用。当地习惯于水煎应用, 可用于产后恢复等, 如产后服用崇明苦草煮鸡蛋, 有助于排出恶露, 降低产后感染等。研究表明, 从妊娠结束到产后恢复, 可能受巨噬细胞激活状态的影响, 与其中IL-6等炎症相关蛋白表达水平有关[2-4]。为此, 本文对崇明苦草水提物中黄酮类成分进行了系统的分离、纯化, 从中得到21个黄酮苷类成分, 其中1个为新化合物, 12个为该种中首次报道。同时采用ELISA法检测了化合物1在不同浓度下对LPS诱导的RAW264.7细胞中IL-6蛋白表达水平, 结果显示, 化合物1在高(100 μmol·L-1)、中(50 μmol·L-1)、低(25 μmol·L-1) 三种浓度下均能表现出显著的抑制IL-6蛋白的表达, 并呈现浓度依赖性, 表明化合物1可通过抑制IL-6的生成和表达来发挥抗炎作用。本文的研究结果, 不仅丰富了黄花蒿水提取物中黄酮类成分的信息, 也可为阐明黄花蒿抗炎作用的药效物质基础提供一定的依据。
化合物1为黄色粉末, 溶于甲醇。根据高分辨质谱的准分子离子峰m/z: 641.170 4 [M+H]+ (计算值641.171 2, C28H33O17) 和13C NMR数据确定分子式为C28H32O17, 不饱和度为13。在260和354 nm处具有最大吸收的紫外光谱表明化合物具有黄酮醇骨架[5]。红外光谱显示, 在3 320 (OH)、1 653 (α-β不饱和CO) 和1 598 (芳香环) cm-1有吸收峰[6]1H NMR谱(表 1) 显示一组ABX耦合系统的3个芳环质子信号δH 7.70 (1H, d, J = 2.2 Hz, H-2′)、6.95 (1H, d, J = 8.5 Hz, H-5′)、7.58 (1H, dd, J = 8.5, 2.2 Hz, H-6′), 表明存在1, 3, 4-三取代环B[7], 以及A环上的一个芳质子信号δH 6.92 (1H, s, H-8), 另可见2个甲氧基信号在δH 3.91 (3H, s, 6-OCH3)、3.82 (3H, s, 3-OCH3), 2个糖的端基氢信号在δH 5.12 (1H, d, J = 7.3 Hz, H-1″)、4.32 (1H, d, J = 7.0 Hz, H-1‴)。化合物113C NMR显示28个碳信号, 除去黄酮骨架15个碳信号和2个甲氧基碳信号, 剩余11个碳信号分别为1个6碳糖和1个5碳糖的信号。
由HSQC可知2个糖端基碳的信号为δC 102.0、105.4, 结合文献[8]和端基氢耦合常数可知一个为β-D-葡萄糖、一个为β-D-木糖。将化合物1的碳谱数据与已知化合物甲氧基万寿菊素-7-O-β-D-吡喃葡萄糖苷(axillarin-7-O-glucopyranoside) 的数据比较[9], 发现二者较为相似, 进一步分析二者的碳谱数据, 化合物1除多了一个木糖片段外, 其余碳信号与已知化合物的基本一致, 唯一区别在葡萄糖C-6位的化学位移不同, 已知化合物中葡萄糖6位碳信号的化学位移值为δC 60.7, 而化合物1的为δC 69.9, 表明化合物1中的木糖片段是连接在葡萄糖的C-6上。HMBC谱(图 1) 中δH 5.12与C-7 (δC 157.8) 相关, δH 4.32 (H-1‴) 与δC 69.9 (C-6″) 相关, 进一步证实化合物1中木糖与葡萄糖是(1→6) 连接的。化合物1的结构鉴定为甲氧基万寿菊素-7-O-β-D-吡喃木糖-(1→6)-β-D-吡喃葡萄糖苷(axillarin-7-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside)。
20个已知化合物分别鉴定为荭草苷(2)[10]、芹菜素-6-C-β-D-吡喃葡萄糖-8-C-β-L-吡喃阿拉伯糖苷(3)[11]、芹菜素-6-C-β-D-吡喃半乳糖-8-C-β-L-吡喃阿拉伯糖苷(4)[12]、芹菜素-6-C-β-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(5)[13, 14]、芹菜素-6-C-α-L-呋喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(6)[15]、槲皮素-3-O-β-D-吡喃葡萄糖苷-(1→2)-β-D-吡喃葡萄糖苷(7)[16]、芹菜素-6-C-α-L-吡喃阿拉伯糖-8-C-β-D-吡喃葡萄糖苷(8)[17]、芹菜素-6, 8-二-C-吡喃葡萄糖苷(9)[18]、万寿菊素-7-O-β-D-吡喃葡萄糖苷(10)[19]、木犀草素-6-C-吡喃葡萄糖苷(11)[20]、牡荆素(12)[21]、山柰酚-3-O-β-吡喃半乳糖-(1→2)-β-吡吡喃葡萄糖苷(13)[22]、槲皮素-7-O-β-D-吡喃葡萄糖苷(14)[23]、万寿菊素-3-O-β-D-吡喃葡萄糖苷(15)[24]、7-甲氧基-栎草亭-6-O-β-D-吡喃葡萄糖苷(16)[25]、槲皮素-3-O-β-D-吡喃葡萄糖苷(17)[26]、假荆芥属苷(18)[27]、芦丁(19)[26]、山柰酚-3-O-β-槐糖苷(20)[28]、万寿菊素-3-O-β-D-芦丁糖苷(21)[29]。其中, 化合物24671011131516182021为首次从黄花蒿中分离得到。
采用ELISA法检测化合物1对LPS诱导的RAW264.7细胞IL-6蛋白表达, 结果表明, 化合物1在高(100 μmol·L-1)、中(50 μmol·L-1)、低(25 μmol·L-1) 三个浓度下IL-6均被显著性抑制(图 2), 并呈现浓度依赖性, 表明化合物1可通过抑制IL-6的生成和表达来发挥抗炎作用。
Bruker AVANCE-Ⅲ (400 MHz或600 MHz) 核磁共振波谱仪(德国Bruker公司), PerkinElmer FT-IR红外光谱仪(美国PerkinElmer公司), TU-1901紫外光谱仪(北京普析通用仪器有限责任公司), Autopol VI型旋光仪(美国Rudolph公司), GRACE中压液相色谱仪(美国GRACE公司), Agilent Technologies 1100制备型色谱仪、安捷伦7000 QQQ GC-MS质谱仪(美国安捷伦公司), CAPCELL PAK C18制备色谱柱(日本资生堂), EYELA WATER BATH SB-2000旋转蒸发器、EYELA COOL ACE CA-1111旋蒸冷凝设备(日本东京理化), SHB-Ⅲ循环水式多用真空泵(郑州长城科工贸有限公司), 电子天平(Sartorius BT124, artorius-BT-323S, 德国赛多利斯集团), MCI填料(75~150 μm)、大孔吸附树脂(HP-20型) (日本三菱化学有限公司), 柱色谱硅胶、薄层色谱及制备型薄层色谱硅胶板(100~200目, 200~300目, 青岛海洋化工厂分厂), Sephadex LH-20 (25~100 μm, 美国通用电器医疗集团), 氘代甲醇、水、吡啶和DMSO (国药集团), 乙腈(制备级和色谱级)、甲醇(制备级、色谱级、分析级)、乙酸乙酯(分析级)、乙醇(分析级)、DMSO (分析级)、吡啶(分析级)、硫酸、甲酸、丙酮(色谱级试剂来源于Fisher公司, 其他均来源于国药公司)。实验用药材购买自上海崇明岛地区, 原植物经上海中医药大学侴桂新研究员鉴定为菊科黄花蒿Artemisia annua L. (20190920-1), 凭证标本保存于上海中药标准化中心。
取崇明苦草32.5 kg, 切段(5~10 mm), 第一次加10倍量水煎煮1 h, 过滤, 第二次药渣加8倍量水煎煮1 h, 过滤, 第三次药渣再加8倍量水煎煮1 h, 过滤, 共提取3次, 合并滤液浓缩至约40 L (60 ℃), 得到崇明苦草药材水提物组分, 编号KC。再将崇明苦草水提物转移至萃取罐中, 按1∶1加入乙酸乙酯, 搅拌, 静置, 分别萃取3次, 合并3次的乙酸乙酯萃取部分, 浓缩干燥得到乙酸乙酯部位, 编号KC-A; 乙酸乙酯萃取剩余部分, 按1∶1加入正丁醇分别萃取3次, 合并3次正丁醇萃取部分, 浓缩干燥得到正丁醇部位, 编号KC-B; 经正丁醇萃取后剩下的水部分, 浓缩干燥得到水部位, 编号KC-C。KC-B部位经HP-20大孔吸附树脂柱分离, 分别依次用水、15%乙醇、30%乙醇、50%乙醇和95%乙醇洗脱, 得到5个部位(KC-B-1~KC-B-5)。取KC-B-3部位经MCI柱色谱分离, 分别依次用30%、40%、50%、60%、100%甲醇洗脱, 得到9个部位(KC-B-301~KC-B-309)。KC-B-301部位进一步采用中压ODS柱色谱分离, 分别用10%、20%、30%、40%、50%、100%甲醇洗脱, 得到15个部位(KC-B-301-1~KC-B-301-15)。
KC-B-301-9部位进一步采用硅胶柱色谱分离, 分别用乙酸乙酯、乙酸乙酯∶甲醇∶水(30∶2∶1、20∶2∶1、15∶2∶1、10∶2∶1)、纯甲醇洗脱, 得到15个部位(KC-B-301-9-1~KC-B-301-9-15)。KC-B-301-9-2部位经液相制备色谱分离(梯度洗脱, 0~30 min, 15%~30%乙腈, 18 min出峰) 可得到化合物2 (2 mg)。KC-B-301-9-5部位凝胶柱色谱分离用10%、20%、30%、40%、50%、60%、70%、80%、90%、100%甲醇洗脱得到14个部位(KC-B-301-9-5-1~KC-B-301-9-5-14), KC-B-301-9-5-10经液相制备色谱分离(洗脱条件为18%乙腈, 14 min出峰) 可得到化合物3 (10 mg), KC-B-301-9-5-11经液相制备色谱分离(梯度洗脱, 0~35 min, 6%~20%乙腈, 31 min出峰) 可得到化合物4 (17 mg), KC-B-301-9-5-12经液相制备色谱分离(洗脱条件为15%乙腈, 26、31 min出峰) 可得到化合物5 (69 mg)、化合物6 (23 mg), KC-B-301-9-5-13经液相制备色谱分离(洗脱条件为20%乙腈, 12 min出峰) 可得到化合物7 (74 mg)。KC-B-301-9-9部位凝胶柱色谱分离用30%、40%、50%、60%、70%、80%、90%、100%的甲醇洗脱得到7个部位(KC-B-301-9-9-1~KC-B-301-9-9-7), KC-B-301-9-9-6部位经液相制备色谱分离(洗脱条件为15%乙腈) 得到化合物8 (37 mg), KC-B-301-9-9-4部位经液相制备色谱分离(洗脱条件为15%乙腈) 得到化合物9 (8 mg)。KC-B-301-12部位用Sephadex LH-20柱色谱分离, 依次用50%醇、70%甲醇、80%甲醇、90%甲醇、无水甲醇洗脱, 得到19个部位(KC-B-301-12-1~KC-B-301-12-19), KC-B-301-12-16部位经液相制备色谱分离可得到化合物10 (18 mg), KC-B-301-12-15部位经液相制备色谱分离(洗脱条件为20%乙腈, 分别在11.5、18.5 min出峰) 可得到化合物11 (3 mg)、12 (20 mg), KC-B-301-12-11部位经液相制备色谱分离(洗脱条件为18%乙腈, 24 min出峰) 可得到化合物13 (30 mg), KC-B-301-12-18部位经液相制备色谱分离可得到化合物14 (12 mg)。KC-B-301-14部位采用硅胶柱色谱分离, 分别用乙酸乙酯∶甲醇∶水(30∶1∶1、20∶1∶1、10∶2∶1)、纯甲醇, 得到7个部位(KC-B-301-14-1~KC-B-301-14-7)。KC-B-301-14-2部位进一步用iSephadex LH-20柱色谱分离, 用40%甲醇洗脱得到11个部位(KC-B-301-14-2-1~KC-B-301-14-2-11), KC-B-301-14-2-8部位经液相制备色谱分离(洗脱条件为20%乙腈) 可得到化合物15 (111 mg), KC-B-301-14-2-10部位经液相制备色谱分离(洗脱条件为20%乙腈) 可得到化合物16 (6 mg)、17 (11 mg)。KC-B-301-14-3部位进一步用Sephadex LH-20柱色谱分离, 用10%、20%、30%、40%、50%、60%、70%、80%的甲醇洗脱得到12个部位(KC-B-301-14-3-1~KC-B-301-14-3-12), KC-B-301-14-3-9部位经液相制备色谱分离(洗脱条件为20%乙腈, 25 min出峰) 可得到化合物18 (25 mg)。KC-B-301-14-4部位用Sephadex LH-20柱色谱分离, 用30%、40%、50%、60%、70%、80%、90%、100%甲醇洗脱得到7个部位(KC-B-301-14-4-1~KC-B-301-14-4-7), KC-B-301-14-4-6部位经液相制备色谱分离(洗脱条件为20%乙腈, 16 min出峰) 可得到化合物19 (800 mg), KC-B-301-14-4-5部位经液相制备色谱分离(梯度洗脱10~30 min, 10%~25%乙腈, 分别在28.8、30.3 min出峰) 可得到化合物20 (12 mg)、21 (12 mg)。KC-B-301-14-4-4部位经液相制备色谱分离(梯度洗脱0~25 min, 15%~40%乙腈, 18 min出峰) 可得到化合物1 (8 mg)。
化合物1: 黄色粉末, HR-ESI-MS m/z: 641.170 4[M+H]+ (计算值C28H32O17, 641.171 2); 旋光值为[α]$ {}_{\mathrm{D}}^{20} $ -47 (c 0.1, MeOH); IR (KBr) υmax/cm-1: 3 321、2 920、1 653、1 598、1 465、1 350、1 212、1 033、805; 1H NMR (400 MHz, methanol-d4) 和13C NMR (100 MHz, methanol-d4) 数据见表 1
将RAW264.7细胞按每毫升1×106的密度接种于6孔板中, 设置空白对照组、LPS (100 ng·mL-1) 模型组、阳性组(槲皮素) 和给药组(高浓度100 μmol·L-1、中浓度50 μmol·L-1、低浓度25 μmol·L-1)。待细胞贴壁后(24 h), 加入一定量稀释好的药物和阳性药, 其终浓度为100、50、25 μmol·L-1, 药物与细胞共同预孵, 孵育2 h后, 加入LPS (100 ng·mL-1) 共同孵育24 h。收集各组每孔培养基上清液用于检测培养基上清中炎症因子分泌量。采用SPSS 21.0统计软件进行分析, 以One-way ANOVA方式进行方差分析, 两两比较采用LSD法, P < 0.05为具有统计学显著性差异标准。结果显示化合物1在高、中、低3个浓度下IL-6均被显著性抑制。
作者贡献: 吴其国是本文第一作者, 负责化合物分离、鉴定、新化合物活性筛选和文章撰写; 范明慧、黄乐怡、王永丽对提取分离提供了一定帮助; 侴桂新为本文通讯作者。
利益冲突: 本文不存在任何利益冲突。
  • 十三五科技重大专项“中药组分资源库”子课题-中药组分及对照品制备(2019ZX09735002-004)
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doi: 10.16438/j.0513-4870.2022-1340
  • 接收时间:2022-12-07
  • 首发时间:2025-11-21
  • 出版时间:2023-06-12
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  • 收稿日期:2022-12-07
  • 修回日期:2022-12-30
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十三五科技重大专项“中药组分资源库”子课题-中药组分及对照品制备(2019ZX09735002-004)
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    1.上海中医药大学中药研究所, 上海 201203
    2.上海中药标准化研究中心, 上海 201203
    3.安庆医药高等专科学校药学院, 安徽 安庆 246052

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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