Article(id=1210147891398709936, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1210147879319113875, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2021-1735, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1638720000000, receivedDateStr=2021-12-06, revisedDate=1641225600000, revisedDateStr=2022-01-04, acceptedDate=null, acceptedDateStr=null, onlineDate=1766451340770, onlineDateStr=2025-12-23, pubDate=1654963200000, pubDateStr=2022-06-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766451340770, onlineIssueDateStr=2025-12-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766451340770, creator=13701087609, updateTime=1766451340770, updator=13701087609, issue=Issue{id=1210147879319113875, tenantId=1146029695717560320, journalId=1189982191388893191, year='2022', volume='57', issue='6', pageStart='1541', pageEnd='1924', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1766451337890, creator=13701087609, updateTime=1766451466252, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1210148417767084534, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1210147879319113875, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1210148417767084535, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1210147879319113875, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1840, endPage=1844, ext={EN=ArticleExt(id=1210147891826528998, articleId=1210147891398709936, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=A new nucleoside from the Oenothera biennis L., columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

Seven nucleoside compounds were isolated from the Oenothera biennis L. by various chromatographic techniques such as Diaion HP-20, silica gel, Sephadex LH-20, MCI and semi-preparative HPLC. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as 9-(3′-carbonyl methyl)hydroxypurine (1), 1-(3′-carbonyl methyl)purine-6, 8-dione (2), N-methyl-2-pyridone-5-carboxamide (3), uracil (4), uridine (5), thymidine (6) and 2′-Ο-methoxy luridine (7). Compound 1 is a new nucleoside and compounds 2-7 were newly isolated from the Oenothera biennis L. Compounds 1-2 can significantly increase the viability of BEAS-2B cells induced by TGF-β1, showing potent anti-pulmonary fibrosis activity.

, correspAuthors=Wei-sheng FENG, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2022 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, authorCompany=null, fund=null, authors=null, authorsList=Juan-juan LIU, Jing-ke ZHANG, Qin-qin ZHANG, Meng LI, Deng-hui ZHU, Jun-jun WEI, Xiao-ke ZHENG, Wei-sheng FENG), CN=ArticleExt(id=1210147892577309510, articleId=1210147891398709936, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=月见草中一个新的核苷类化合物, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

运用大孔吸附树脂、硅胶、凝胶Sephadex LH-20、MCI等柱色谱并结合半制备高效液相等分离纯化技术, 从月见草中分离得到7个核苷类化合物, 根据化合物的理化性质和波谱特征对其结构进行鉴定, 分别为9-(3′-carbonyl methyl)hydroxypurine (1)、1-(3′-carbonyl methyl)purine-6, 8-dione (2)、N-methyl-2-pyridone-5-carboxamide (3)、尿嘧啶(4)、尿嘧啶核苷(5)、胸腺嘧啶脱氧核苷(6) 和2′-Ο-甲氧基尿嘧啶核苷(7)。其中化合物1为新化合物, 化合物2~7为首次从月见草中分离得到。化合物12可显著提高TGF-β1诱导BEAS-2B细胞的活力, 说明其具有潜在的抗肺纤维化活性。

, correspAuthors=冯卫生, authorNote=null, correspAuthorsNote=
*冯卫生, Tel: 86-371-60190296, E-mail:
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No. δH δC
1 12.3 (1H, brs) -
2 8.04 (1H, brs) 145.5
3 - -
4 - 148.2
5 - 123.6
6 - 156.5
7 - -
8 8.06 (1H, s) 140.4
9 - -
1′ 4.30 (2H, t, J = 6.8 Hz) 38.4
2′ 3.08 (2H, t, J = 6.8 Hz) 42.2
3′ - 206.2
4′ 2.10 (3H, s) 29.8
), ArticleFig(id=1210147898365448559, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1210147891398709936, language=CN, label=Table 1, caption=

1H NMR (500 MHz, DMSO-d6) and 13C NMR (125 MHz, DMSO-d6) spectral data of compound 1

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No. δH δC
1 12.3 (1H, brs) -
2 8.04 (1H, brs) 145.5
3 - -
4 - 148.2
5 - 123.6
6 - 156.5
7 - -
8 8.06 (1H, s) 140.4
9 - -
1′ 4.30 (2H, t, J = 6.8 Hz) 38.4
2′ 3.08 (2H, t, J = 6.8 Hz) 42.2
3′ - 206.2
4′ 2.10 (3H, s) 29.8
), ArticleFig(id=1210147898457723256, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1210147891398709936, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
Group Cell index
Con 1.000 ± 0.009**
M 0.835 ± 0.068
1 0.945 ± 0.020*
2 0.946 ± 0.025**
3 0.833 ± 0.012
4 0.861 ± 0.073
5 0.891 ± 0.031
6 0.912 ± 0.036
7 0.905 ± 0.032
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The effect of compounds 1-7 on BEAS-2B cell injury induced by TGF-β1 (x±s, n = 4). Con: Normal group; M: Model group. *P < 0.05; **P < 0.01 vs M group

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Group Cell index
Con 1.000 ± 0.009**
M 0.835 ± 0.068
1 0.945 ± 0.020*
2 0.946 ± 0.025**
3 0.833 ± 0.012
4 0.861 ± 0.073
5 0.891 ± 0.031
6 0.912 ± 0.036
7 0.905 ± 0.032
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月见草中一个新的核苷类化合物
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刘娟娟 1, 2 , 张靖柯 1, 2 , 张钦钦 1, 2 , 李孟 1, 2 , 朱登辉 1, 2 , 魏俊俊 1, 2 , 郑晓珂 1, 2 , 冯卫生 1, 2, *
药学学报 | 研究论文 2022,57(6): 1840-1844
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药学学报 | 研究论文 2022, 57(6): 1840-1844
月见草中一个新的核苷类化合物
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刘娟娟1, 2, 张靖柯1, 2, 张钦钦1, 2, 李孟1, 2, 朱登辉1, 2, 魏俊俊1, 2, 郑晓珂1, 2, 冯卫生1, 2, *
作者信息
  • 1.河南中医药大学药学院, 河南 郑州 450046
  • 2.河南省中药开发工程技术研究中心, 河南 郑州 450046

通讯作者:

*冯卫生, Tel: 86-371-60190296, E-mail:
A new nucleoside from the Oenothera biennis L.
Juan-juan LIU1, 2, Jing-ke ZHANG1, 2, Qin-qin ZHANG1, 2, Meng LI1, 2, Deng-hui ZHU1, 2, Jun-jun WEI1, 2, Xiao-ke ZHENG1, 2, Wei-sheng FENG1, 2, *
Affiliations
  • 1. College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China
  • 2. The Engineering and Technology Center for Chinese Medicine Development of Henan Province, Zhengzhou 450046, China
出版时间: 2022-06-12 doi: 10.16438/j.0513-4870.2021-1735
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运用大孔吸附树脂、硅胶、凝胶Sephadex LH-20、MCI等柱色谱并结合半制备高效液相等分离纯化技术, 从月见草中分离得到7个核苷类化合物, 根据化合物的理化性质和波谱特征对其结构进行鉴定, 分别为9-(3′-carbonyl methyl)hydroxypurine (1)、1-(3′-carbonyl methyl)purine-6, 8-dione (2)、N-methyl-2-pyridone-5-carboxamide (3)、尿嘧啶(4)、尿嘧啶核苷(5)、胸腺嘧啶脱氧核苷(6) 和2′-Ο-甲氧基尿嘧啶核苷(7)。其中化合物1为新化合物, 化合物2~7为首次从月见草中分离得到。化合物12可显著提高TGF-β1诱导BEAS-2B细胞的活力, 说明其具有潜在的抗肺纤维化活性。

月见草  /  化学成分  /  核苷类  /  BEAS-2B细胞  /  MTT

Seven nucleoside compounds were isolated from the Oenothera biennis L. by various chromatographic techniques such as Diaion HP-20, silica gel, Sephadex LH-20, MCI and semi-preparative HPLC. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as 9-(3′-carbonyl methyl)hydroxypurine (1), 1-(3′-carbonyl methyl)purine-6, 8-dione (2), N-methyl-2-pyridone-5-carboxamide (3), uracil (4), uridine (5), thymidine (6) and 2′-Ο-methoxy luridine (7). Compound 1 is a new nucleoside and compounds 2-7 were newly isolated from the Oenothera biennis L. Compounds 1-2 can significantly increase the viability of BEAS-2B cells induced by TGF-β1, showing potent anti-pulmonary fibrosis activity.

Oenothera biennis L.  /  chemical constituent  /  nucleoside  /  BEAS-2B cell  /  MTT
刘娟娟, 张靖柯, 张钦钦, 李孟, 朱登辉, 魏俊俊, 郑晓珂, 冯卫生. 月见草中一个新的核苷类化合物. 药学学报, 2022 , 57 (6) : 1840 -1844 . DOI: 10.16438/j.0513-4870.2021-1735
Juan-juan LIU, Jing-ke ZHANG, Qin-qin ZHANG, Meng LI, Deng-hui ZHU, Jun-jun WEI, Xiao-ke ZHENG, Wei-sheng FENG. A new nucleoside from the Oenothera biennis L.[J]. Acta Pharmaceutica Sinica, 2022 , 57 (6) : 1840 -1844 . DOI: 10.16438/j.0513-4870.2021-1735
月见草Oenothera biennis L.为柳叶菜科月见草属多年生草本植物, 最早出自《贵州草药》, 全草入药[1]。性温、味甘, 具有强筋壮骨、祛风除湿、活血通络的功效。可用于治疗胸痹心痛, 中风偏瘫, 虚风内动, 小儿多动等症状[2]。现代药理研究表明, 月见草具有抗菌[3]、抗炎[4]、抗氧化[5]、降血脂[6, 7]、降血糖[8, 9]、抗肿瘤[10]等药理作用。原产于北美洲, 在我国主要产于东北南部及东部山区等地。其中在河南伏牛山区的山坡、荒地、河岸等地随处可见, 资源分布十分丰富。目前国内外对于月见草的研究主要集中在其种子上, 而对月见草的化学成分缺乏系统的研究。为了进一步对月见草化学成分进行系统的研究, 为其后续药效物质基础研究提供理论依据, 本实验利用大孔树脂、硅胶、ODS、Sephadex LH-20、MCI等柱色谱并结合半制备高效液相等分离纯化技术, 从月见草50%含水丙酮提取物中分离得到7个核苷类化合物, 根据化合物的理化性质及波谱特征对其结构进行鉴定。分别为9-(3′-carbonyl methyl)hydroxypurine (1)、1-(3′-carbonyl methyl)purine-6, 8-dione (2)、N-methyl-2-pyridone-5-carboxamide (3)、尿嘧啶(4)、尿嘧啶核苷(5)、胸腺嘧啶脱氧核苷(6) 和2′-Ο-甲氧基尿嘧啶核苷(7)。其中化合物1为新化合物, 化合物2~7为首次从月见草中分离得到, 化合物的结构见图 1
肺纤维化(pulmonary fibrosis, PF) 是一种慢性、不可逆性和致死性的肺间质病[11], 研究表明, 多种呼吸道病毒(包括常见的流感以及突发的新冠病毒) 感染均可能导致肺纤维化的发生[12]。目前, 对于该病的治疗尚无效果显著的药物, 临床常用的激素、尼达尼布等都存在一定的不良反应[13]。中药具有悠久的历史, 其作用广泛、不良反应少、药物依赖性低等, 且越来越多的研究表明中药及其提取物对肺纤维化有治疗作用[14-16]。本研究采用TGF-β1诱导BEAS-2B细胞建立肺纤维化体外模型, 基于MTT法检测化合物1~7对TGF-β1诱导BEAS-2B细胞损伤的影响。结果表明, 化合物12可显著提高TGF-β1诱导BEAS-2B细胞的活力, 说明其具有潜在的抗肺纤维化活性。
化合物1, 白色粉末, [α]D20 -3.062 (c 0.080 4, CH3OH), HR-ESI-MS m/z: 207.087 7 [M+H]+ (C9H11N4O2计算值207.088 2), 提示化合物的分子式为C9H10N4O2。UV λmax (CH3OH)/nm (logε): 203 (1.90)、249 (0.96), IR光谱中3 358 cm-1为伯胺N-H伸缩振动特征峰, 1 640 cm-1有C=N双键伸缩振动特征峰。
1H NMR (500 MHz, DMSO-d6) 谱中, δH 8.06 (1H, s, H-8)、8.04 (1H, brs, H-2) 推测为氮杂环上不饱和氢的信号; δH 12.3 (1H, brs, -NH) 为一个活泼氢的信号; δH 4.30 (2H, t, J = 6.8 Hz, H-1′)、3.08 (2H, t, J = 6.8 Hz, H-2′) 为两个亚甲基的氢信号; δH 2.10 (3H, s, H-4′) 为一个甲基的氢信号。在13C NMR (125 MHz, DMSO-d6) 谱中, 共有9个碳信号, 结合DEPT135谱可知, 共有4个季碳信号[δC 123.6 (C-5)、148.2 (C-4)、156.5 (C-6)、206.2 (C-3′), 其中δC 156.5 (C-6)[17]、206.2 (C-3′) 为两个酮羰基的碳信号], 2个叔碳信号[δC 140.4 (C-8)、145.5 (C-2)], 2个仲碳信号[δC 38.4 (C-1′)、42.2 (C-2′)], 1个伯碳信号[δC 29.8 (C-4′)]。
1H-1H COSY谱中, δH 4.30 (2H, t, J = 6.8 Hz, H-1′) 与3.08 (2H, t, J = 6.8 Hz, H-2′) 相关, 确定-CH2-CH2-的结构片段。通过HSQC谱, 进一步将化合物的C-H数据予以归属(表 1)。HMBC谱中δH 2.10 (3H, s, H-4′) 与δC 38.4 (C-1′)、42.2 (C-2′)、206.2 (C-3′) 具有远程相关, 可以确定-CH2CH2COCH3的结构片段; δH 8.04 (1H, brs, H-2) 与δC 148.2 (C-4)、123.6 (C-5)、156.5 (C-6) 具有远程相关, 且δH 8.06 (1H, brs, H-8) 与δC 148.2 (C-4)、123.6 (C-5) 也具有远程相关, 说明存在一个次黄嘌呤的结构片段; 同时根据δH 4.30 (2H, t, J = 6.8 Hz, H-1′) 与δC 148.2 (C-4)、140.4 (C-8) 具有远程相关, 说明-CH2CH2COCH3连在次黄嘌呤9位N原子上(图 2)。根据以上数据, 结合文献NMR数据[18], 与文献中已知化合物相比差别仅在于C-4′位酮羰基, 确定为新化合物, 结构为9-(3′-carbonyl methyl)hydroxypurine。
采用TGF-β1诱导BEAS-2B细胞建立肺纤维化体外模型, 基于MTT法检测化合物1~7对TGF-β1诱导BEAS-2B细胞损伤的影响。结果表明, 化合物12可显著提高TGF-β1诱导BEAS-2B细胞的活力, 提示其可能具有潜在的抗肺纤维化活性(表 2)。
本研究从月见草中分离并鉴定了7个核苷类化合物, 其中化合物1为新化合物, 化合物2~7为首次从月见草中分离得到。化合物12可显著提高TGF-β1诱导BEAS-2B细胞的活力, 说明其具有潜在的抗肺纤维化活性, 为月见草进一步的研究与开发提供了一定的理论依据。
Bruker AVANCE Ⅲ 500型核磁共振仪(德国Bruker公司); Bruker maxis HD型飞行时间质谱(德国Bruker公司); OSB-21000型旋转蒸发仪(上海爱朗仪器有限公司); 赛谱锐思LC-52型高压制备液相色谱仪(赛谱锐思北京科技有限公司, SP-5030型半制备型高压输液泵, UV200型紫外检测器, Easychrom型色谱工作站; 色谱柱为COSMOSIL C18-MS-Ⅱ, 250 mm × 20 mm, 5 μm); 薄层色谱硅胶GF 254; Sephadex LH-20 (Pharmacia Biotech公司); 柱色谱所用硅胶H (200~300目) 为青岛海洋化工厂生产; 所用色谱纯试剂为天津市富宇精细化工有限公司生产; 所用分析纯试剂为北京化工厂和天津第三化学试剂厂生产; 二氧化碳3111型培养箱(Thermo, USA); Multiskan MK3酶标仪(Thermo Fisher, USA); Advantage A10超纯水仪(Sartorius, Germany); ECTGF-Β1E TS100倒置显微镜(Nikon, Japan); TGF-β1 (PeproTech, 美国); 噻唑蓝(北京索莱宝科技有限公司); 二甲基亚砜(上海麦克林生物科技有限公司)。
月见草采自于河南省伏牛山区, 经河南中医药大学董诚明教授鉴定为柳叶菜科月见草属植物月见草(Oenothera biennis L.) 的干燥全草。
取干燥月见草40 kg, 用50%含水丙酮组织破碎提取2次, 过滤, 合并滤液, 减压浓缩, 得到总浸膏3.3 kg。总浸膏加等量水分散, 依次用石油醚、二氯甲烷、乙酸乙酯以及水饱和正丁醇进行萃取, 各部位经减压浓缩, 回收溶剂, 得到石油醚部位(5.0 g)、二氯甲烷部位(49.6 g)、乙酸乙酯部位(161.0 g) 及正丁醇部位(457.0 g)。对正丁醇部位上Diaion HP-20大孔吸附树脂柱, 按照极性由大到小依次用水、10% EtOH、20% EtOH、30% EtOH、50% EtOH、70% EtOH、95% EtOH进行梯度洗脱, 得到7个洗脱组分, 即Fr.1~Fr.7。
Fr.2 (52.0 g) 加水溶解, 进行凝胶Sephadex LH-20柱色谱, 用不同比例的甲醇-水(甲醇∶水= 0∶100→100∶0) 梯度洗脱, 洗脱组分经薄层硅胶检识, 合并相同组分, 得到6个洗脱组分: Fr.2-1~Fr.2-6。其中Fr.2-2经凝胶Sephadex LH-20柱色谱, 70%含水甲醇等度洗脱, 经硅胶薄层检识, 合并相同组分, 得到4个洗脱组分, 即Fr.2-2-1~Fr.2-2-4, Fr.2-2-2反复经过凝胶Sephadex LH-20柱色谱分离并结合半制备高效液相(乙腈-水洗脱体系) 最终得到化合物1 (4.02 mg)、化合物5 (12.15 mg) 和化合物7 (3.25 mg); Fr.2-2-4加适量水溶解, 进行MCI柱色谱分离, 用不同比例的甲醇-水(甲醇∶水= 0∶100→100∶0) 梯度洗脱, 最终得到4个洗脱组分, 即Fr.2-2-4-1~Fr.2-2-4-4, Fr.2-2-4-2进行制备薄层分离纯化, 以二氯甲烷-甲醇-水系统为展开剂(二氯甲烷∶甲醇∶水= 10∶1∶0.1), 最终得到化合物4 (5.56 mg)、化合物6 (8.89 mg); Fr.2-3采用Fr.2-2-2相同的洗脱方法, 得到3个洗脱组分, 即Fr.2-3-1~Fr.2-3-3, Fr.2-3-3加适量甲醇溶解, 硅胶(200~300目) 拌样后进行硅胶柱色谱分离, 用二氯甲烷-甲醇系统(二氯甲烷∶甲醇= 30∶1) 洗脱, 得到3个洗脱组分Fr.2-3-3-1~Fr.2-3-3-3, Fr.2-3-3-2使用半制备高效液相(乙腈-水洗脱体系) 最终得到化合物2 (9.84 mg); Fr.2-4加适量水溶解, 进行ODS柱色谱分离, 用不同比例的甲醇-水(甲醇∶水= 0∶100→100∶0) 梯度洗脱, 最终得到5个洗脱组分, 即Fr.2-4-1~Fr.2-4-5, Fr.2-4-1反复经过Sephadex LH-20、Toyopearl HW-40凝胶柱色谱分离, 结合使用半制备高效液相(乙腈-水洗脱体系), 最终得到化合物3 (6.35 mg)。
化合物1  白色粉末, 溶于甲醇, HR-ESI-MS m/z: 207.087 7 [M+H]+ (C9H10N4O2计算值207.088 2), 提示化合物的分子式为C9H10N4O2。UV λmax (CH3OH)/nm (logε): 203 (1.90)、249 (0.96), IR (CH3OH) vmax: 3 358、1 640 cm-11H NMR (500 MHz, DMSO-d6) 和13C NMR (125 MHz, DMSO-d6) 数据见表 1
化合物2  白色粉末, 溶于甲醇, ESI-MS m/z: 223 [M+H]+, 分子式为C9H10N4O31H NMR (500 MHz, DMSO-d6) δH 7.88 (1H, s, H-2), 4.32 (2H, t, J = 6.6 Hz, H-1′), 3.06 (2H, t, J = 6.6 Hz, H-2′), 2.09 (3H, s, H-4′); 13C NMR (125 MHz, DMSO-d6) δC 142.8 (C-2), 149.5 (C-4), 106.1 (C-5), 155.5 (C-6), 151.2 (C-8), 41.1 (C-1′), 43.0 (C-2′), 206.2 (C-3′), 29.8 (C-4′)。根据以上数据, 结合参考文献[17], 鉴定该化合物为1-(3′-carbonylbutyl) purine-6, 8-dione。
化合物3  白色粉末, 溶于甲醇, ESI-MS m/z: 153 [M+H]+, 分子式为C7H8N2O21H NMR (500 MHz, DMSO-d6) δH 8.46 (1H, d, J = 2.5 Hz, H-6), 7.77 (1H, dd, J = 2.5, 9.4 Hz, H-4), 6.40 (1H, d, J = 9.4 Hz, H-3), 3.49 (3H, s, H-7); 13C NMR (125 MHz, DMSO-d6) δC 162.0 (C-2), 118.1 (C-3), 138.8 (C-4), 109.0 (C-5), 144.9 (C-6), 37.3 (C-7), 165.4 (C-8)。根据以上数据, 结合参考文献[19], 确定化合物为N-methyl-2-pyridone-5-carboxamide。
化合物4  白色无定形粉末, 溶于甲醇, ESI-MS m/z: 135 [M+Na]+, 分子式为C4H4N2O21H NMR (500 MHz, DMSO-d6) δH 11.01 (1H, s, NH-3), 10.81 (1H, s, NH-1), 7.38 (1H, d, J = 7.7 Hz, H-6), 5.44 (1H, d, J = 7.7 Hz, H-5); 13C NMR (125 MHz, DMSO-d6) δC 151.5 (C-2), 164.4 (C-4), 100.2 (C-5), 142.2 (C-6)。根据以上数据, 结合参考文献[20], 确定化合物为尿嘧啶。
化合物5  白色针晶, 溶于甲醇, ESI-MS m/z: 267 [M+Na]+, 分子式为C9H12N2O61H NMR (500 MHz, DMSO-d6) δH 8.01 (1H, d, J = 8.1 Hz, H-6), 5.90 (1H, d, J = 4.6 Hz, H-1′), 5.69 (1H, d, J = 8.1 Hz, H-5), 4.16 (2H, m, H-2′), 4.00 (1H, m, H-3′), 3.84 (1H, m, H-4′), 3.73 (1H, m, H-5′); 13C NMR (125 MHz, DMSO-d6) δC 152.4 (C-2), 166.2 (C-4), 102.6 (C-5), 142.7 (C-6), 90.7 (C-1′), 71.3 (C-2′), 75.7 (C-3′), 86.4 (C-4′), 62.3 (C-5′)。根据以上数据, 结合参考文献[21], 确定化合物为尿嘧啶核苷。
化合物6  白色粉末, 溶于甲醇, ESI-MS m/z: 265 [M+Na]+, 分子式为C10H14N2O51H NMR (500 MHz, DMSO-d6) δH 11.25 (1H, s, -NH), 7.69 (1H, s, H-6), 6.16 (1H, t, J = 6.3 Hz, H-1′), 4.23 (1H, m, H-3′), 3.75 (1H, m, H-4′), 3.56 (2H, m, H-5′), 2.07 (2H, m, H-2′), 1.76 (3H, s, -CH3); 13C NMR (125 MHz, DMSO-d6) δC 150.5 (C-2), 163.7 (C-4), 109.4 (C-5), 136.1 (C-6), 83.7 (C-1′), 39.4 (C-2′), 70.4 (C-3′), 87.2 (C-4′), 61.3 (C-5′), 12.31 (-CH3)。根据以上数据, 结合参考文献[22], 确定化合物为胸腺嘧啶脱氧核苷。
化合物7  白色粉末, 溶于甲醇, ESI-MS m/z: 281 [M+Na]+, 分子式为C10H14N2O61H NMR (500 MHz, DMSO-d6) δH 8.08 (1H, d, J = 8.1 Hz, H-6), 5.94 (1H, d, J = 3.6 Hz, H-1′), 5.68 (1H, d, J = 8.1 Hz, H-5), 4.33 (1H, t, J = 6.0 Hz, H-3′), 3.97 (1H, m, H-4′), 3.86 (2H, m, H-5′), 3.74 (1H, dd, J = 3.0, 12.4 Hz, H-2′), 3.52 (3H, s, 2′-OCH3); 13C NMR (125 MHz, DMSO-d6) δC 152.2 (C-2), 166.2 (C-4), 102.5 (C-5), 142.4 (C-6), 88.8 (C-1′), 85.0 (C-2′), 69.8 (C-3′), 86.1 (C-4′), 61.6 (C-5′), 58.8 (2′-OCH3)。根据以上数据, 结合参考文献[23], 确定化合物为2′-Ο-甲氧基尿嘧啶核苷。
采用MTT法探究7个化合物对TGF-β1诱导BEAS-2B细胞损伤的影响[24]。将BEAS-2B细胞置于37 ℃、5% CO2培养箱中培养至对数生长期, 按照细胞密度每毫升2.5×104个细胞, 每孔200 μL接种于96孔板中, 24 h后分为正常组(Control)、模型组(TGF-β1, 1 ng·mL-1) 及各给药组(10 μmol·L-1 + TGF-β1, 1 ng·mL-1), 继续培养24 h后, 每孔加入MTT溶液(5 mg·mL-1) 20 μL, 再继续培养4 h, 小心吸净培养液, 每孔加入DMSO 150 μL, 振荡10 min使孔内蓝紫色结晶完全溶解。酶标仪490 nm测定各孔吸光度值(OD), 计算细胞活力。实验数据以均值±标准差(x±s) 表示, 应用SPSS 26.0进行统计学分析, 以P < 0.05为差异有统计学意义。
作者贡献: 刘娟娟主要负责月见草化学成分的提取、分离、化合物结构鉴定和文章的整理; 张靖柯对月见草化学成分提取、分离、结构鉴定以及文章写作时提供帮助; 张钦钦对从月见草中分离得到的化合物进行活性筛选; 李孟对月见草化学成分分离以及化合物结构鉴定出现困难时进行指导; 朱登辉、魏俊俊、郑晓柯对月见草的文献查找、化学成分提取、分离提供帮助; 冯卫生为通讯作者, 对稿件负总责。
利益冲突: 本文不存在利益冲突。
  • 河南省重大科技专项(171100310500)
  • 国家重点研发计划-中医药现代化研究(2019YFC1708802)
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2022年第57卷第6期
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doi: 10.16438/j.0513-4870.2021-1735
  • 接收时间:2021-12-06
  • 首发时间:2025-12-23
  • 出版时间:2022-06-12
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  • 收稿日期:2021-12-06
  • 修回日期:2022-01-04
基金
河南省重大科技专项(171100310500)
国家重点研发计划-中医药现代化研究(2019YFC1708802)
作者信息
    1.河南中医药大学药学院, 河南 郑州 450046
    2.河南省中药开发工程技术研究中心, 河南 郑州 450046

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*冯卫生, Tel: 86-371-60190296, E-mail:
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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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