Article(id=1228268593847136998, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, articleNumber=null, orderNo=null, doi=10.12048/j.issn.1674-229X.2025.08.003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1722960000000, receivedDateStr=2024-08-07, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1770771652900, onlineDateStr=2026-02-11, pubDate=1756051200000, pubDateStr=2025-08-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770771652900, onlineIssueDateStr=2026-02-11, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770771652900, creator=13701087609, updateTime=1770771652900, updator=13701087609, issue=Issue{id=1228268587459216114, tenantId=1146029695717560320, journalId=1227999776411738115, year='2025', volume='35', issue='8', pageStart='561', pageEnd='640', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1770771651377, creator=13701087609, updateTime=1770795163631, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1228367205105075039, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1228367205105075040, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=576, endPage=582, ext={EN=ArticleExt(id=1228268594132349693, articleId=1228268593847136998, tenantId=1146029695717560320, journalId=1227999776411738115, language=EN, title=A Comparative Study on the Antibacterial, Antiviral, and Anti-Inflammatory Activities of Sea Buckthorn Leaf Extracts from Different Origins, columnId=null, journalTitle=Pharmacy Today, columnName=null, runingTitle=null, highlight=null, articleAbstract=
OBJECTIVE

To compare the antibacterial (against Escherichia coli/Salmonella), antiviral (against HSV-1/2), and NF-κB inhibitory activity of sea buckthorn leaf extracts (SBLE) from different origins.

METHODS

SBLEs were prepared using 65% ethanol for hot reflux extraction of sea buckthorn leaves from different origins. The differences in their fingerprint profiles were analyzed by liquid chromatography. The colorimetric method was used to detect the inhibitory activity of Escherichia coli and Salmonella. The CCK-8 assay was employed to evaluate the activity of Vero cells infected with HSV-1/2. The luciferase reporter assay was conducted to assess the activity of NF-κB in TNF-α-induced NF-κB reporter (Luc)-HEK 293 cells.

RESULTS

SBLE from Datong and Huzhu exhibited better inhibitory effects on Escherichia coli (with EC50 values of 184.3 and 211.6 μg·mL-1, respectively) and Salmonella (with EC50 values of 284.6 and 300.9 μg·mL-1, respectively). SBLE from Datong, Pingan, Huzhu, Youyu, Heshun, and Zhangbei showed similar inhibitory effects on HSV-1 and HSV-2, with EC50 values ranging from 27 to 36 μg·mL-1. SBLE from Heshun, Zhangbei, and Huzhu exhibited better inhibition of NF-κB, with EC50 values of 122.6, 137.1, and 154.1 μg·mL-1, respectively.

CONCLUSION

The activities of SBLE from different origins vary, with SBLE from Datong exhibiting the best anti-Escherichia coli/Salmonella activity, SBLE from Youyu showing the best anti-HSV-1/2 activity, and SBLE from Heshun having the best NF-κB inhibitory effect.

, correspAuthors=Lei LIANG, Yifang LI, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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=Xiaolun ZHANG, Zhaojun DING, Changyu YAN, Xiaomin LI, Jian ZOU, Zhengchao TU, Zhen WANG, Kurihara HIROSHI, Rongrong HE, Lei LIANG, Yifang LI), CN=ArticleExt(id=1228268597747839889, articleId=1228268593847136998, tenantId=1146029695717560320, journalId=1227999776411738115, language=CN, title=不同产地沙棘叶提取物抗菌、抗病毒与抗炎活性的比较, columnId=1228268589086606089, journalTitle=今日药学, columnName=论著, runingTitle=null, highlight=null, articleAbstract=
目的

比较不同产地沙棘叶提取物(sea buckthorn leaves extracts,SBLE)的抗大肠杆菌/沙门氏菌、抗HSV-1/2病毒与NF-κB抑制作用。

方法

65%乙醇对不同产地的沙棘叶进行热回流提取制备SBLE,液相色谱分析其指纹图谱的差异。比色法检测大肠杆菌和沙门氏菌活性,CCK-8实验检测HSV-1/2感染Vero细胞的活力。荧光素酶报告实验检测TNF-α诱导NF-κB reporter(Luc)-HEK 293细胞中NF-κB的活性。

结果

大通与互助产地SBLE对大肠杆菌(EC50分别为184.3和211.6 μg·mL-1)和沙门氏菌(EC50分别为284.6和300.9 μg·mL-1)的抑制作用较好。大通、平安、互助、右玉、和顺和张北6个产地的SBLE对HSV-1和HSV-2的抑制作用相近,EC50在27~36 μg·mL-1范围内。和顺、张北、互助产地的SBLE对NF-κB的抑制效果较好,EC50分别为122.6、137.1和154.1 μg·mL-1

结论

不同产地SBLE的活性存在差异,大通产地SBLE的抗大肠杆菌/沙门氏菌活性最好,右玉产地SBLE的抗HSV-1/2病毒活性最好,和顺产地SBLE的NF-κB抑制作用最好。

, correspAuthors=梁磊, 李怡芳, authorNote=null, correspAuthorsNote=
* 李怡芳,博士,教授,博士生导师,研究方向:中药药理,E-mail:
梁磊,博士,研究员,博士生导师,研究方向:中医病因病机和中药药理,E-mail:
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张孝伦,研究方向:中药药理学

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张孝伦,研究方向:中药药理学

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张孝伦,研究方向:中药药理学

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Biomed Pharmacother, 2023(158): 114199., articleTitle=Therapeutic effects of traditional Chinese medicine on gouty nephropathy: Based on NF-κB signalingpathways, refAbstract=null), Reference(id=1228268610674684161, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, doi=null, pmid=null, pmcid=null, year=2024, volume=41, issue=12, pageStart=1663, pageEnd=1670, url=null, language=null, rfNumber=[26], rfOrder=25, authorNames=江望, 田生望, 贺单, journalName=中国现代应用药学, refType=null, unstructuredReference=江望, 田生望, 贺单, 等. 三仁汤对幽门螺杆菌相关性胃炎脾胃湿热证大鼠消化吸收功能及Akt/NF-κB通路的影响[J]. 中国现代应用药学, 2024, 41(12): 1663-1670., articleTitle=三仁汤对幽门螺杆菌相关性胃炎脾胃湿热证大鼠消化吸收功能及Akt/NF-κB通路的影响, refAbstract=null)], funds=[Fund(id=1228268606992085190, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, awardId=2023LSYS002, language=CN, fundingSource=广东省粤港澳高校联合实验室项目(2023LSYS002), fundOrder=null, country=null), Fund(id=1228268607075971272, 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journalId=1227999776411738115, articleId=1228268593847136998, language=CN, label=图2, caption=不同产地SBLE的指纹图谱

注:A.不同提取溶剂制备SBLE(吕梁)的指纹图谱;B.不同产地SBLE的指纹图谱。

, figureFileSmall=4rPwiGtevoNk64cisPMIww==, figureFileBig=TU+jkSphq2kceD3DOPl/Xw==, tableContent=null), ArticleFig(id=1228268606367133878, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=EN, label=null, caption=null, figureFileSmall=RUGiFREmQvoRW41sXFW3Cg==, figureFileBig=m38dtFrSFmTwqncBmdQ7CA==, tableContent=null), ArticleFig(id=1228268606434242745, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=CN, label=图3, caption=不同产地SBLE对大肠杆菌和沙门氏菌的影响(n=3)

注:A. SBLE对大肠杆菌的影响;B. SBLE对沙门氏菌的影响。

, figureFileSmall=RUGiFREmQvoRW41sXFW3Cg==, figureFileBig=m38dtFrSFmTwqncBmdQ7CA==, tableContent=null), ArticleFig(id=1228268606530711740, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=EN, label=null, caption=null, figureFileSmall=oTRBPWfN6Y+jsR3wTHwF2w==, figureFileBig=Gt00Ko8wOJgho8bqjhLEZg==, tableContent=null), ArticleFig(id=1228268606614597823, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=CN, label=图4, caption=不同产地SBLE对HSV-1/2病毒的影响(n=3)

注:A. SBLE对Vero细胞活力的影响;B. SBLE对HSV-1病毒的影响;C. SBLE对HSV-2病毒的影响。

, figureFileSmall=oTRBPWfN6Y+jsR3wTHwF2w==, figureFileBig=Gt00Ko8wOJgho8bqjhLEZg==, tableContent=null), ArticleFig(id=1228268606702678209, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=EN, label=null, caption=null, figureFileSmall=/g/IgGJxLFLr3JyVgcihzg==, figureFileBig=kvraoAtCGUzQCOjc8SlU2g==, tableContent=null), ArticleFig(id=1228268606883033282, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268593847136998, language=CN, label=图5, caption=不同产地SBLE的NF-κB抑制活性(n=3)

注:A. SBLE对HEK293细胞活力的影响;B. SBLE对NF-κB活性的影响。

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不同产地沙棘叶提取物抗菌、抗病毒与抗炎活性的比较
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张孝伦 1, 2 , 丁肇俊 2, 3 , 闫昌誉 2, 3 , 李晓敏 4, 5 , 邹剑 2, 3 , 涂正超 3 , 王珍 3 , 栗原博 2, 3, 4, 5 , 何蓉蓉 2, 3 , 梁磊 1, 2, 3, * , 李怡芳 2, 3, *
今日药学 | 论著 2025,35(8): 576-582
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今日药学 | 论著 2025, 35(8): 576-582
不同产地沙棘叶提取物抗菌、抗病毒与抗炎活性的比较
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张孝伦1, 2, 丁肇俊2, 3, 闫昌誉2, 3, 李晓敏4, 5, 邹剑2, 3, 涂正超3, 王珍3, 栗原博2, 3, 4, 5, 何蓉蓉2, 3, 梁磊1, 2, 3, * , 李怡芳2, 3, *
作者信息
  • 1.暨南大学中医学院,广东 广州 510632
  • 2.暨南大学中药与天然药物研究所,广东 广州 510632
  • 3.暨南大学药学院,广东 广州 510632
  • 4.完美(广东)日用品有限公司,广东 中山 528400
  • 5.广东完美生命健康科技研究院有限公司,广东 中山 528400
  • 张孝伦,研究方向:中药药理学

通讯作者:

* 李怡芳,博士,教授,博士生导师,研究方向:中药药理,E-mail:
梁磊,博士,研究员,博士生导师,研究方向:中医病因病机和中药药理,E-mail:
A Comparative Study on the Antibacterial, Antiviral, and Anti-Inflammatory Activities of Sea Buckthorn Leaf Extracts from Different Origins
Xiaolun ZHANG1, 2, Zhaojun DING2, 3, Changyu YAN2, 3, Xiaomin LI4, 5, Jian ZOU2, 3, Zhengchao TU3, Zhen WANG3, Kurihara HIROSHI2, 3, 4, 5, Rongrong HE2, 3, Lei LIANG1, 2, 3, * , Yifang LI2, 3, *
Affiliations
  • 1.School of Traditional Chinese Medicine, Jinan University, Guangzhou, Guangdong 510632, China
  • 2.Institute of Traditional Chinese Medicine and Natural Products, Jinan University, Guangzhou, Guangdong 510632, China
  • 3.School of Pharmacy, Jinan University, Guangzhou, Guangdong 510632, China
  • 4.Perfect (Guangdong) Commodity Co., Ltd., Zhongshan, Guangdong 528400, China
  • 5.Perfect Life Sciences Research Institute Co., Ltd., Zhongshan, Guangdong 528400, China
出版时间: 2025-08-25 doi: 10.12048/j.issn.1674-229X.2025.08.003
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目的

比较不同产地沙棘叶提取物(sea buckthorn leaves extracts,SBLE)的抗大肠杆菌/沙门氏菌、抗HSV-1/2病毒与NF-κB抑制作用。

方法

65%乙醇对不同产地的沙棘叶进行热回流提取制备SBLE,液相色谱分析其指纹图谱的差异。比色法检测大肠杆菌和沙门氏菌活性,CCK-8实验检测HSV-1/2感染Vero细胞的活力。荧光素酶报告实验检测TNF-α诱导NF-κB reporter(Luc)-HEK 293细胞中NF-κB的活性。

结果

大通与互助产地SBLE对大肠杆菌(EC50分别为184.3和211.6 μg·mL-1)和沙门氏菌(EC50分别为284.6和300.9 μg·mL-1)的抑制作用较好。大通、平安、互助、右玉、和顺和张北6个产地的SBLE对HSV-1和HSV-2的抑制作用相近,EC50在27~36 μg·mL-1范围内。和顺、张北、互助产地的SBLE对NF-κB的抑制效果较好,EC50分别为122.6、137.1和154.1 μg·mL-1

结论

不同产地SBLE的活性存在差异,大通产地SBLE的抗大肠杆菌/沙门氏菌活性最好,右玉产地SBLE的抗HSV-1/2病毒活性最好,和顺产地SBLE的NF-κB抑制作用最好。

沙棘叶  /  活性比较  /  大肠杆菌  /  沙门氏菌  /  HSV-1/2  /  NF-κB
OBJECTIVE

To compare the antibacterial (against Escherichia coli/Salmonella), antiviral (against HSV-1/2), and NF-κB inhibitory activity of sea buckthorn leaf extracts (SBLE) from different origins.

METHODS

SBLEs were prepared using 65% ethanol for hot reflux extraction of sea buckthorn leaves from different origins. The differences in their fingerprint profiles were analyzed by liquid chromatography. The colorimetric method was used to detect the inhibitory activity of Escherichia coli and Salmonella. The CCK-8 assay was employed to evaluate the activity of Vero cells infected with HSV-1/2. The luciferase reporter assay was conducted to assess the activity of NF-κB in TNF-α-induced NF-κB reporter (Luc)-HEK 293 cells.

RESULTS

SBLE from Datong and Huzhu exhibited better inhibitory effects on Escherichia coli (with EC50 values of 184.3 and 211.6 μg·mL-1, respectively) and Salmonella (with EC50 values of 284.6 and 300.9 μg·mL-1, respectively). SBLE from Datong, Pingan, Huzhu, Youyu, Heshun, and Zhangbei showed similar inhibitory effects on HSV-1 and HSV-2, with EC50 values ranging from 27 to 36 μg·mL-1. SBLE from Heshun, Zhangbei, and Huzhu exhibited better inhibition of NF-κB, with EC50 values of 122.6, 137.1, and 154.1 μg·mL-1, respectively.

CONCLUSION

The activities of SBLE from different origins vary, with SBLE from Datong exhibiting the best anti-Escherichia coli/Salmonella activity, SBLE from Youyu showing the best anti-HSV-1/2 activity, and SBLE from Heshun having the best NF-κB inhibitory effect.

sea buckthorn leaves  /  activity comparison  /  Escherichia coli  /  Salmonella  /  HSV-1/2  /  NF-κB
张孝伦, 丁肇俊, 闫昌誉, 李晓敏, 邹剑, 涂正超, 王珍, 栗原博, 何蓉蓉, 梁磊, 李怡芳. 不同产地沙棘叶提取物抗菌、抗病毒与抗炎活性的比较. 今日药学, 2025 , 35 (8) : 576 -582 . DOI: 10.12048/j.issn.1674-229X.2025.08.003
Xiaolun ZHANG, Zhaojun DING, Changyu YAN, Xiaomin LI, Jian ZOU, Zhengchao TU, Zhen WANG, Kurihara HIROSHI, Rongrong HE, Lei LIANG, Yifang LI. A Comparative Study on the Antibacterial, Antiviral, and Anti-Inflammatory Activities of Sea Buckthorn Leaf Extracts from Different Origins[J]. Pharmacy Today, 2025 , 35 (8) : 576 -582 . DOI: 10.12048/j.issn.1674-229X.2025.08.003
沙棘(Hippophae rhamnoides L.)是一种雌雄异株、落叶、带刺的灌木或小乔木,天然分布于亚洲和欧洲等地区,而我国拥有世界上90%的沙棘资源[1]。因其耐旱、耐盐碱、对土壤营养要求低、防风固沙等特点,一直作为沙漠绿化植物在种植。沙棘具有独特的风味,其果实在食品领域被广泛应用,被开发为沙棘果汁、沙棘茶等多种产品[2]。沙棘亦有一定的药用价值,在我国传统医药中常用于利肺化痰、止咳平喘、活血化瘀等[3]。而现代药理学研究显示沙棘具有抗氧化、免疫调节、抗癌、保肝和降血脂等生物功能[4,5]。沙棘果实、叶子、根皮和种子等部位均含有丰富的营养物质和生物活性成分[6]。其中,沙棘叶具有易采摘保存、蕴藏量巨大、黄酮含量更为丰富等优势[7]。2013年,国家卫生和计划生育委员会第7号公告将沙棘叶作为新资源食品进行管理,这极大地拓宽了沙棘叶的应用市场。我国国土面积广袤,地理形势复杂多样,海拔、气候、降雨、土壤等因素会影响的沙棘的生长与成熟。因此,不同地区沙棘叶的活性成分和及其含量可能存在差异,这将直接影响沙棘叶的功效。然而,目前的研究主要集中于部分地区的沙棘叶,而对不同产地沙棘叶的比较分析却鲜有报道。本研究对产自7个地区沙棘叶提取物(sea buckthorn leaves extracts,SBLE)的抑菌、抗病毒、抗炎活性进行了比较分析,以期为沙棘叶相关功能产品的研发提供指导。
非洲绿猴肾细胞(Vero细胞)由中国科学院广州生物医药科学与健康研究院馈赠,培养于含5%FBS、1%P/S的DMEM培养基。NF-κB reporter(Luc)-HEK 293细胞(Cat. 60650)购于BPS Bioscience公司,培养于含10%FBS、1%P/S、1 mmol·L-1丙酮酸钠、1%NEAA、50 μg·mL-1 Hygromycin B的MEM培养基。大肠埃希氏菌(Lot. CICC10305)和乙型副伤寒沙门氏菌(Lot. CICC21495)购于中国工业微生物菌种保藏管理中心。HSV-1(F株)和HSV-2(333株)由暨南大学药学院药物筛选中心库存。
7个产地的沙棘叶(图1)由完美(广东)日用品有限公司提供,经暨南大学岭南传统中药研究中心张英老师鉴定为胡颓子科植物沙棘(Hippophae rhamnoides L.)的干燥叶,分别产自平安(青海,Lot.20200805)、互助(青海,Lot. 20200805)、大通(青海,Lot. 20200805)、和顺(山西,Lot. 20200809)、右玉(山西,Lot. 20200809)、吕梁(山西,Lot. 20200809)、张北(河北,Lot. 20200809)。
无水乙醇(分析级)购自天津大茂化学试剂厂;甲醇(色谱级)、乙腈(色谱级)购自德国Merck公司;环丙沙星(Cat. L1711047)购自上海Aladdin公司;阿昔洛韦(Cat. A4669)、IKK-16(Cat. SML1138)、TNF-α(Cat. T0157)购自美国Sigma公司;CCK-8(Cat.B34304)购自美国Bimake公司;DMEM培养基(Cat.C11995500BT)、MEM培养基(Cat. SH30024.01)购自美国Thermo公司;Reporter lysis 5x buffer(Cat.E397A)、Luciferase assay substrate(Cat. E151A)购自美国Promega公司。
Echo 550超微量液体移液系统(美国Labcyte公司);AC2-5S1Ⅱ级A2型生物安全柜、CCL-170B-8细胞培养箱(新加坡ESCO公司);Envision多模式读板仪(美国PerkinElmer公司);AS 220.R2分析天平(波兰Radwag公司);Ultimate 3000高效液相色谱仪、Multiskan Mk 3酶标仪(美国Thermo公司);D-16C高速冷冻离心机、PB-10精密pH调节计(德国Sartorius公司);Milli Q超级纯水仪(美国Millipore公司);SB-5200D超声波清洗机(宁波新芝生物);DHP-9032电热恒温培养箱(上海一恒仪器)。
称取30 g沙棘叶,置于1 L圆底烧瓶,分别加入95%乙醇、65%乙醇和纯水进行热回流提取2 h,重复提取2次,合并提取液后抽滤、减压浓缩、转移、称重,计算收率。
称取2 mg的SBLE,加入1 mL 50%色谱甲醇超声溶解,14 000 r·min-1离心20 min,吸取上清过0.22 μm微孔滤膜得到浓度为2 mg·mL-1的样品。分析仪器为Ultimate 3000 DGLC液相色谱仪,色谱柱为Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8 μm)。流动相为0.1%甲酸水(A)-乙腈(B),洗脱程序如下,0 min:5%(B),1 min:12%(B),10 min:16%(B),14 min:32%(B),15 min:100%(A),18.5 min:100%(B),19 min:5%(B),25 min:5%(B),流速为0.3 mL·min-1,柱温为40 ℃,紫外扫描波长为254 nm,进样量为2 μL。
将活化的大肠杆菌或沙门氏菌以1∶3 000稀释,接种于384孔板中,每孔25 μL菌液。使用Echo handler 550移液器向菌液中加入梯度稀释的SBLE样品,放置于37 ℃培养箱孵育16 h,不含菌液的空白培养基中转移同样浓度的样品作为药物对照组,不含样品的菌液作为菌液对照组,培养基作为空白对照组,环丙沙星(Ciprofloxacin)作为阳性对照药。培养结束后于570 nm检测吸光度,并根据下列公式计算,细菌抑制率(%)=[1-(Abs药物处理组 -Abs药物对照组)/(Abs细菌对照组 -Abs空白对照组)]×100。
将Vero细胞或HEK293细胞种板于384孔板,5 000个/孔。细胞过夜培养后,利用Echo handler 550移液器转移梯度稀释的沙棘叶样品至细胞板中,37 ℃培养箱孵育72 h,不含细胞的空白培养基中转移同样浓度的样品作为药物对照组,不含样品的细胞作为细胞对照组,培养基作为空白对照组。去除培养基加入1∶20稀释的CCK-8溶液,每孔15 μL。37 °C孵育2 h,采用酶标仪检测450 nm处吸收值,根据下列公式计算,细胞活力(%)=[(Abs药物处理组 -Abs药物对照组)/(Abs细胞对照组 -Abs空白对照组)]×100。
Vero细胞接种于384孔板中,细胞过夜培养后,利用Echo handler 550移液器转移梯度稀释的沙棘叶样品至细胞板中,37 ℃培养箱孵育30 min,阿昔洛韦(Acyclovir)作为阳性对照。用HSV-1病毒感染细胞,HSV-1(F株)稀释比例为1∶70 000,HSV-2(333株)稀释比例为1∶5 000,将细胞置于37 ℃培养箱继续培养72 h,不加样品的感染细胞作为病毒对照组,不加样品且未感染的细胞作为细胞对照组。去除培养基,加入CCK-8进行细胞活力检测,于450 nm处检测吸光度。根据下列公式计算,病毒抑制率(%)=[(Abs药物处理组 -Abs病毒对照组)/(Abs细胞对照组 -Abs病毒对照组)]×100。
NF-κB reporter(Luc)-HEK 293细胞接种于384孔板中,过夜培养后利用Echo handler 550移液器转移梯度稀释的沙棘叶样品至细胞板中,37 ℃孵育2 h,IKK-16作为阳性对照。接着加入TNF-α(10 ng·mL-1)诱导NF-κB激活,37 ℃箱孵育5 h。去除培养基,采用Luciferase Assay System试剂盒进行萤火虫萤光素酶检测:加入Reporter lysis buffer裂解细胞,震荡15 min后离心;转移5 μL细胞裂解液上清,加入5 μL luciferase assay substrate反应5 min后检测化学发光。根据下列公式计算,NF-κB抑制率(%)=[1-(Luc药物处理组-LucnoTNF-α)/(LucTNF-α-LucnoTNF-α)]×100。
使用GraphPad Prism 8.0进行统计分析,计量资料以均数±标准差()表示。
为了优化SBLE的制备条件,笔者采用纯水、95%乙醇与65%乙醇作为溶剂对沙棘叶(吕梁)进行热回流提取,收率分别为19.20%、9.03%和24.90%,可见65%乙醇提取获得了更高的收率。此外,还通过色谱比较了3种SBLE活性成分的溶出,结果如图2A所示。沙棘叶65%乙醇提取物的色谱吸收峰相较纯水和95%乙醇更为丰富。因此,笔者后续采用65%乙醇制备其它产地的SBLE。结果显示,来自平安和互助的SBLE收率最高,分别为41.23%和40.13%,其余产地的收率为33.80%(大通)、35.47%(和顺)、38.57%(右玉)、24.90%(吕梁)和37.70%(张北)。各产地SBLE的液相分析显示,7个产地提取物的主要吸收峰保留时间基本一致,提示其化学成分相似度较高,但仍有部分吸收峰有所不同。此外,主要吸收峰的峰面积也不相同,提示其化学成分的含量存在差异(图2B)。
本研究计算了各产地SBLE体外抑制大肠杆菌(Escherichia coli)和沙门氏菌(Salmonella)的半数有效剂量EC50。结果如图3A所示,SBLE对大肠杆菌的抑制作用强弱顺序为大通>平安>互助>右玉>和顺,张北和吕梁产地的SBLE抑制作用最弱(EC50 > 300 μg·mL-1)。对于沙门氏菌,仅有大通与互助产地的SBLE具有一定的抑制作用,其EC50分别为284.60、300.90 μg·mL-1,而其余产地的抑制效果较差(图3B)。阳性药环丙沙星抑制大肠杆菌和沙门氏菌的EC50分别为2.97×10-6、4.44×10-6 μg·mL-1图3)。
Vero细胞是一种广泛应用于病毒学领域的工具细胞系,可用于病毒的扩增、疫苗的制备和病毒学研究等[8]。本文选用Vero细胞评价SBLE体外的抗HSV病毒活性。首先,评价了SBLE对Vero细胞活力的影响,发现各产地SBLE在100~300 μg·mL-1浓度范围内对Vero细胞均有一定的毒性作用(图4A)。随后,在安全剂量范围内评价了各地SBLE(除吕梁外)对HSV-1(F株)和HSV-2(333株)两种病毒株的抗病毒效果。6个产地的SBLE对HSV-1(F株)和HSV-2(333株)均有抑制作用,且效果相近,抑制病毒的EC50在27~36 μg·mL-1范围内。阳性药阿昔洛韦抑制HSV-1(F株)和HSV-2(333株)的EC50分别为4.39×10-8 μg·mL-1和5.54×10-8 μg·mL-1,结果见图4BC
细胞在受到促炎刺激后,NF-κB转录因子会向核内转移并与相应DNA响应元件结合,启动多种炎症因子的转录。reporter(Luc)-HEK293重组细胞中NF-κB的响应元件与荧光素酶的基因串联,因而荧光素酶的表达可用于监测NF-κB的活化水平[9]。本文评估了SBLE对NF-κB reporter(Luc)-HEK293细胞活力的影响,结果显示各产地SBLE(除吕梁外)在100 μg·mL-1浓度范围内对细胞具有较好的安全性(图5A),后续在此浓度范围内进行抗炎活性评价。TNF-α是一种常见的炎性因子,可与细胞膜表面的TNF-α受体结合促进下游NF-κB的激活[10],本研究在此模型上比较了不同产地SBLE的抗炎活性。和顺、张北、互助、大通(EC50 <200 μg·mL-1)SBLE对TNF-α诱导NF-κB的抑制活性优于平安和右玉(EC50 >200 μg·mL-1),其中和顺活性最强,EC50值为122.6 μg·mL-1图5B)。阳性药IKK-16是IKK的选择性抑制剂,可通过抑制IKK复合物而影响下游NF-κB的活性[11],其抑制NF-κB活化的EC50为1.56×10-7 μg·mL-1图5B)。
沙棘叶中含有丰富的活性物质,具有广泛的药理作用[12,13],前期研究表明,沙棘叶醇提物在体内外均具有良好的降糖作用[14]。因此,沙棘叶作为一种重要的药用资源,其资源利用的最大化与最优化也是值得关注的问题。笔者在制备SBLE的过程中发现,相较于水提,醇提法能够更好地促进提取物的溶出,而65%的乙醇不仅能够获得更高的收率,且提取物中的成分更加丰富。在此提取条件下,本文比较了7个产地沙棘叶的提取率,发现平安与互助地区的沙棘叶具有较高的收率,对资源的利用更充分。通过高效液相色谱法制备的指纹图谱比较发现,7个产地SBLE的吸收峰虽然大致相同,但仍存在差异,这些化学成分的差异仍需要进一步通过质谱技术进行定性和定量分析。这种差异可能与产地的气候、土壤条件等有关,有研究者利用最大熵(Maximum Entropy,MaxEnt)模型对中国沙棘自然分布的328个地理样点的主导气候因子分析,发现水分条件是影响中国沙棘种群分布的主要气候因子[15]。由于本实验属于初步探索,未对指纹图谱进行方法学验证,色谱分离结果有待进一步提升,但对于初步得出上述结论没有影响,后续将在深入研究中进行改善提高。
致病菌污染引起的食品安全事故仍是危害人民健康的重大公共卫生问题,常见的食源性致病菌包括大肠杆菌、沙门菌、金黄色葡萄球菌、副溶血性弧菌、蜡样芽孢杆菌等[16]。之前的研究表明SBLE具有较广的抗菌谱,对革兰阳性菌与革兰阴性菌、以及一些厌氧菌均有一定的抑制作用,这与其含有的黄酮、多酚、挥发油等活性组分有关[17]。本文发现大通、平安、互助、右玉、和顺5个产地的SBLE对大肠杆菌有抑制作用(EC50在150~300 μg·mL-1范围内);大通与互助2个产地SBLE对沙门氏菌同样有抑制活性,其EC50分别为284.60 μg·mL-1和300.90 μg·mL-1。此外,先前的研究也表明,SBLE发挥抑菌活性的浓度需达到100 μg·mL-1以上[18,19]。因此,选择适宜产地的沙棘叶制备提取物,有望添加到食品中以减缓食源性病原体的生长,从而延长食品的保质期。
疱疹病毒(Herpes simplex virus,HSV)是一类DNA病毒,属疱疹病毒科(Herpesviridae),主要包括2个亚型,即单纯疱疹病毒类型1(HSV-1)和单纯疱疹病毒类型2(HSV-2),广泛分布于全球[20]。HSV-1通常与口唇周围的感染有关,是引起口唇疱疹的常见原因。HSV-2主要与生殖器疱疹相关,这两种类型的疱疹病毒都能在人体内潜伏,并在某些情况下被激活复发,导致反复的感染症状[21,22]。有关沙棘叶抗病毒作用的报道较少,Enkhtaivan等[23]研究发现,沙棘叶甲醇和乙酸乙酯提取物对甲型流感病毒具有抑制活性,其EC50分别为7.2 μg·mL-1和10.3 μg·mL-1。本研究结果表明,大通、平安、互助、右玉、和顺和张北6个产地的SBLE对HSV-1和HSV-2表现出相近的抗病毒活性,EC50在27~36 μg·mL-1范围内。沙棘叶作为食品原料,其提取物是用于开发相关产品的直接形式,相对环丙沙星与阿昔洛韦等药物,SBLE含有多种天然活性成分,不易产生耐药性,且可长期食用,具有较好的安全性,在食品领域具有应用优势。
细菌与病毒感染通常会引发炎症反应,导致组织与器官的炎性损伤。其中,NF-κB通路在感染过程中可被激活,促进各种炎症相关基因的表达,是炎症反应的关键介导者[24]。因此,NF-κB常作为炎性疾病的干预靶点,研究表明许多中药都具有显著的NF-κB抑制与抗炎活性[25,26]。在本研究中,通过TNF-α诱导的炎症模型,比较了6个不同产地SBLE对NF-κB的抑制作用。结果显示作用强弱顺序为和顺>张北>互助>大通>平安>右玉,提示不同产地的SBLE在抗炎活性方面存在差异。
综上所述,不同产地SBLE的抗菌、抗病毒与抗炎活性存在差异,大通产地的SBLE具有最佳的抗大肠杆菌/沙门氏菌活性,右玉产地的SBLE表现出最强的抗HSV-1/2病毒活性,和顺产地的SBLE在NF-κB抑制作用方面效果最佳。因此,在制备SBLE时,应根据所需功能选取合适产地的沙棘叶原料,以实现最优的功效。实际上,不同产地SBLE的差异决定了其功效的不同,后续应进一步解析生物活性关联的化学成分,以揭示其功效差异的物质基础并发掘新的化合物,以期开发出更好的抗菌、抗病毒产品,进一步充分利用沙棘叶资源。
  • 广东省粤港澳高校联合实验室项目(2023LSYS002)
  • 广东省创新团队项目(2020KCXTD003)
  • 暨南大学国家级大学生创新创业训练计划资助(202410559092)
  • 中山市院士工作站建设项目(中山科发[2019]187号)
  • 中山市第八批创新创业科研团队项目(CTXD2020003)
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2025年第35卷第8期
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doi: 10.12048/j.issn.1674-229X.2025.08.003
  • 接收时间:2024-08-07
  • 首发时间:2026-02-11
  • 出版时间:2025-08-25
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  • 收稿日期:2024-08-07
基金
广东省粤港澳高校联合实验室项目(2023LSYS002)
广东省创新团队项目(2020KCXTD003)
暨南大学国家级大学生创新创业训练计划资助(202410559092)
中山市院士工作站建设项目(中山科发[2019]187号)
中山市第八批创新创业科研团队项目(CTXD2020003)
作者信息
    1.暨南大学中医学院,广东 广州 510632
    2.暨南大学中药与天然药物研究所,广东 广州 510632
    3.暨南大学药学院,广东 广州 510632
    4.完美(广东)日用品有限公司,广东 中山 528400
    5.广东完美生命健康科技研究院有限公司,广东 中山 528400

通讯作者:

* 李怡芳,博士,教授,博士生导师,研究方向:中药药理,E-mail:
梁磊,博士,研究员,博士生导师,研究方向:中医病因病机和中药药理,E-mail:
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https://castjournals.cast.org.cn/joweb/jryx/CN/10.12048/j.issn.1674-229X.2025.08.003
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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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