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Article(id=1308842004738564457, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308841897058198067, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2026-0112, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1769443200000, receivedDateStr=2026-01-27, revisedDate=1775318400000, revisedDateStr=2026-04-05, acceptedDate=null, acceptedDateStr=null, onlineDate=1789981851038, onlineDateStr=2026-09-21, pubDate=1783785600000, pubDateStr=2026-07-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789981851038, onlineIssueDateStr=2026-09-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789981851038, creator=13701087609, updateTime=1789981851038, updator=13701087609, issue=Issue{id=1308841897058198067, tenantId=1146029695717560320, journalId=1189982191388893191, year='2026', volume='61', issue='7', pageStart='2377', pageEnd='2820', issueExtLink='null', onlineDate='null', pubDate='1783785600000', pubDateStr='2026-07-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789981825365, creator='13701087609', updateTime=1789984154811, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1308851667525792021, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308841897058198067, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1308851667525792022, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308841897058198067, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2596, endPage=2602, ext={EN=ArticleExt(id=1308842004931502442, articleId=1308842004738564457, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Two new lanostane-type triterpenoids with antidepressant activity from Ganoderma lucidum, columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

Two compounds were isolated from the traditional Chinese medicine Ganoderma lucidum using various chromatographic techniques. Through methods such as one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy, both compounds were identified as new lanostane-type triterpenoids. Their absolute configurations were determined via quantum chemical calculations, and they were named baoslingzhines X and Y (1 and 2). In vitro and in vivo experimental results revealed that these two compounds exhibit potent activity against neuroinflammation, oxidative stress, and depression. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Shenzhen University (approval number: IACUC-202400148).

, authors=Fan FANG1, Yun-yun LIU2, Zi ZHU2, Wei-fen LI2, Yong-xian CHENG2, *, authorsList=Fan FANG, Yun-yun LIU, Zi ZHU, Wei-fen LI, Yong-xian CHENG, authorCompany=null, correspAuthors=Yong-xian CHENG, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2026 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=1308842008102396281, articleId=1308842004738564457, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=赤芝中2个具有抗抑郁活性的新的羊毛脂甾烷型三萜的分离与鉴定, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

利用多种色谱技术从中药赤芝中分离得到2个化合物。经一维(1D)、二维(2D)核磁共振波谱(NMR)等方法鉴定其均为新的羊毛脂甾烷型三萜, 并采用量子化学计算方法确定其绝对构型, 命名为baoslingzhines X和Y (1和2)。体内外实验结果发现, 这两个化合物具有较强的抗神经炎症、抗氧化应激和抑郁症的活性。所有动物实验均获得深圳大学动物伦理与福利委员会批准(批准号: IACUC-202400148)。

, authors=方凡1, 刘云云2, 朱紫2, 李惟芬2, 程永现2, *, authorsList=方凡, 刘云云, 朱紫, 李惟芬, 程永现, authorCompany=null, correspAuthors=程永现, authorNote=

作者贡献:

方凡和朱紫是本文的第一作者和第三作者, 负责生物活性的检测; 刘云云为本文的第二作者, 负责结构鉴定; 李惟芬为本文的第四作者, 负责数据的处理和文章的撰写; 程永现为本文的通讯作者, 负责实验的指导和设计, 并对文章进行审阅和定稿。

, correspAuthorsNote=
程永现, Tel: 86-755-86172799, E-mail:
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NC: The normal control group, no treatment; LPS: The model group, treated with the inflammatory inducer LPS; LPS+1/2: the experimental groups treated with compounds 1 or 2; LPS + Fluoxetine: The positive control group. A: Body weight; B: Sucrose preference test; C: Tail suspension test; D: Forced swimming test. n = 5, x ± s. *P < 0.05, ***P < 0.001, ****P < 0.000 1 vs NC group; #P < 0.05, ##P < 0.01, ####P < 0.000 1 vs LPS group, figureFileSmall=GNPLYRwyxXXDlDz3Ikt5hQ==, figureFileBig=Dnr1J1XBK0AEMPU96ENW9g==, tableContent=null), ArticleFig(id=1308842014452572596, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308842004738564457, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
No.1a2a
δHδCδHδC
1188.2, C3.97, dd (9.4, 7.3)85.0, CH
25.21, s97.9, CHHa: 3.00, dd (12.4, 9.4)42.7, CH2
Hb: 2.69, dd (12.4, 7.3)
3207.5, C215.9, C
445.3, C46.2, C
52.16, dd (14.5, 2.8)50.9, CH2.47, dd (12.7, 3.5)48.7, CH
6Ha: 2.69, dd (16.8, 14.5)36.9, CH2Ha: 2.57, dd (16.6, 12.7)36.7, CH2
Hb: 2.44, dd (16.8, 2.8)Hb: 2.38, m
7200.6, C201.8, C
8138.5, C135.4, C
9156.3, C164.3, C
1044.5, C43.4, C
115.60, dd (11.3, 8.0)83.5, CH4.57, dd (9.0, 7.9)76.7, CH
12Ha: 2.62, dd (11.3, 8.0)34.3, CH2Ha: 2.40, m34.5, CH2
Hb: 1.89, t-like (11.3)Hb: 1.80, m
1350.4, C49.2, C
1449.8, C50.4, C
15Ha: 1.97, t (10.8)30.8, CH2Ha: 1.94, m31.5, CH2
Hb: 1.76, mHb: 1.77, m
16Ha: 2.09, m29.2, CH2Ha: 2.08, m29.5, CH2
Hb: 1.48, mHb: 1.47, m
171.67, m49.5, CH1.63, m49.4, CH
180.98, s17.0, CH30.91, s16.7, CH3
191.50, s18.5, CH31.14, s18.7, CH3
201.58, m37.2, CH1.54, m37.3, CH
211.09, d (6.5)19.3, CH31.06, d (6.3)19.3, CH3
22Ha: 1.63, m36.1, CH2Ha: 1.61, m36.1, CH2
Hb: 1.26, mHb: 1.25, m
23Ha: 2.28, m26.5, CH2Ha: 2.27, m26.5, CH2
Hb: 2.19, mHb: 2.16, m
246.79, t (6.8)144.1, CH6.79, t-like (7.6)144.2, CH
25128.9, C128.8, C
26171.7, C171.7, C
271.82, s12.4, CH31.82, s12.4, CH3
281.21, s21.2, CH31.09, s27.6, CH3
291.10, s29.2, CH31.09, s19.0, CH3
300.96, s26.1, CH30.98, s25.5, CH3
), ArticleFig(id=1308842014523875765, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308842004738564457, language=CN, label=Tab.1, caption=

1H (600 MHz), 13C NMR (150 MHz) data of 1 and 2 (δ in ppm, J in Hz). aRecorded in methanol-d4

, figureFileSmall=null, figureFileBig=null, tableContent=
No.1a2a
δHδCδHδC
1188.2, C3.97, dd (9.4, 7.3)85.0, CH
25.21, s97.9, CHHa: 3.00, dd (12.4, 9.4)42.7, CH2
Hb: 2.69, dd (12.4, 7.3)
3207.5, C215.9, C
445.3, C46.2, C
52.16, dd (14.5, 2.8)50.9, CH2.47, dd (12.7, 3.5)48.7, CH
6Ha: 2.69, dd (16.8, 14.5)36.9, CH2Ha: 2.57, dd (16.6, 12.7)36.7, CH2
Hb: 2.44, dd (16.8, 2.8)Hb: 2.38, m
7200.6, C201.8, C
8138.5, C135.4, C
9156.3, C164.3, C
1044.5, C43.4, C
115.60, dd (11.3, 8.0)83.5, CH4.57, dd (9.0, 7.9)76.7, CH
12Ha: 2.62, dd (11.3, 8.0)34.3, CH2Ha: 2.40, m34.5, CH2
Hb: 1.89, t-like (11.3)Hb: 1.80, m
1350.4, C49.2, C
1449.8, C50.4, C
15Ha: 1.97, t (10.8)30.8, CH2Ha: 1.94, m31.5, CH2
Hb: 1.76, mHb: 1.77, m
16Ha: 2.09, m29.2, CH2Ha: 2.08, m29.5, CH2
Hb: 1.48, mHb: 1.47, m
171.67, m49.5, CH1.63, m49.4, CH
180.98, s17.0, CH30.91, s16.7, CH3
191.50, s18.5, CH31.14, s18.7, CH3
201.58, m37.2, CH1.54, m37.3, CH
211.09, d (6.5)19.3, CH31.06, d (6.3)19.3, CH3
22Ha: 1.63, m36.1, CH2Ha: 1.61, m36.1, CH2
Hb: 1.26, mHb: 1.25, m
23Ha: 2.28, m26.5, CH2Ha: 2.27, m26.5, CH2
Hb: 2.19, mHb: 2.16, m
246.79, t (6.8)144.1, CH6.79, t-like (7.6)144.2, CH
25128.9, C128.8, C
26171.7, C171.7, C
271.82, s12.4, CH31.82, s12.4, CH3
281.21, s21.2, CH31.09, s27.6, CH3
291.10, s29.2, CH31.09, s19.0, CH3
300.96, s26.1, CH30.98, s25.5, CH3
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赤芝中2个具有抗抑郁活性的新的羊毛脂甾烷型三萜的分离与鉴定
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方凡 1 , 刘云云 2 , 朱紫 2 , 李惟芬 2 , 程永现 2, *
药学学报 | 研究论文 2026,61(7): 2596-2602
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药学学报 |研究论文 2026 , 61 (7) : 2596 -2602
赤芝中2个具有抗抑郁活性的新的羊毛脂甾烷型三萜的分离与鉴定
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方凡1, 刘云云2, 朱紫2, 李惟芬2, 程永现2, *
作者信息
  • 1云南中医药大学, 基础医学院, 云南 昆明 650500
  • 2深圳大学医学部药学院, 中医药守正创新研究院, 广东 深圳 518060
通讯作者:
程永现, Tel: 86-755-86172799, E-mail:
作者简介:

作者贡献:

方凡和朱紫是本文的第一作者和第三作者, 负责生物活性的检测; 刘云云为本文的第二作者, 负责结构鉴定; 李惟芬为本文的第四作者, 负责数据的处理和文章的撰写; 程永现为本文的通讯作者, 负责实验的指导和设计, 并对文章进行审阅和定稿。

Two new lanostane-type triterpenoids with antidepressant activity from Ganoderma lucidum
Fan FANG1, Yun-yun LIU2, Zi ZHU2, Wei-fen LI2, Yong-xian CHENG2, *
Affiliations
  • 1School of Basic Medical Sciences, Yunnan University of Chinese Medicine, Kunming 650500, China
  • 2Institute for Inheritance-Based Innovation of Chinese Medicine, Shenzhen University Medical School, Shenzhen University, Shenzhen 518060, China
出版时间: 2026-07-12 doi: 10.16438/j.0513-4870.2026-0112
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摘要
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利用多种色谱技术从中药赤芝中分离得到2个化合物。经一维(1D)、二维(2D)核磁共振波谱(NMR)等方法鉴定其均为新的羊毛脂甾烷型三萜, 并采用量子化学计算方法确定其绝对构型, 命名为baoslingzhines X和Y (1和2)。体内外实验结果发现, 这两个化合物具有较强的抗神经炎症、抗氧化应激和抑郁症的活性。所有动物实验均获得深圳大学动物伦理与福利委员会批准(批准号: IACUC-202400148)。

赤芝  /  分离鉴定  /  羊毛脂甾烷型三萜  /  神经炎症  /  抑郁症

Two compounds were isolated from the traditional Chinese medicine Ganoderma lucidum using various chromatographic techniques. Through methods such as one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy, both compounds were identified as new lanostane-type triterpenoids. Their absolute configurations were determined via quantum chemical calculations, and they were named baoslingzhines X and Y (1 and 2). In vitro and in vivo experimental results revealed that these two compounds exhibit potent activity against neuroinflammation, oxidative stress, and depression. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Shenzhen University (approval number: IACUC-202400148).

Ganoderma lucidum  /  isolation and characterization  /  lanostane-type triterpenoid  /  neuroinflammation  /  depression
方凡, 刘云云, 朱紫, 李惟芬, 程永现. 赤芝中2个具有抗抑郁活性的新的羊毛脂甾烷型三萜的分离与鉴定. 药学学报, 2026 , 61 (7) : 2596 -2602 . DOI: 10.16438/j.0513-4870.2026-0112
Fan FANG, Yun-yun LIU, Zi ZHU, Wei-fen LI, Yong-xian CHENG. Two new lanostane-type triterpenoids with antidepressant activity from Ganoderma lucidum[J]. Acta Pharmaceutica Sinica, 2026 , 61 (7) : 2596 -2602 . DOI: 10.16438/j.0513-4870.2026-0112
正文
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抑郁症(depression)是一种常见的精神心理障碍, 严重影响患者的情绪、行为及生活质量[1]。其核心症状包括持续性情绪低落、丧失对日常活动的兴趣和愉悦性, 以及能量和动机的显著下降[2]。目前的治疗策略多包括药物治疗和心理干预, 但抑郁症患者因药物治疗的不良反应等导致治疗结果不理想[3]。灵芝, 在中国已被广泛使用了数千年, 最早见于《神农本草经》, 性平, 味甘, 有“益心气, 补中, 增慧智, 益脾气, 安神, 利关节, 保神, 益精气, 坚筋骨, 好颜色”等功效[4]。其为灵芝科真菌赤芝[Ganoderma lucidum (Leyss. ex Fr.) Karst.] 或紫芝(Ganoderma sinense Zhao, Xu et Zhang)的干燥子实体, 普遍种植于全国各地[5, 6]。迄今为止, 从灵芝子实体、孢子和菌丝体中提取出多种生物活性代谢产物, 以多糖类和三萜类化合物为主要成分[7, 8], 并具有免疫调节、抗炎、抗氧化、抗肿瘤、神经保护、肝脏保护、降血糖和降血脂等多种药理活性[9]。灵芝作为传统药食两用真菌, 毒性反应轻微, 临床耐受性良好。因此, 针对灵芝“安神”的功效, 深入开展其抗抑郁活性成分分离鉴定与生物活性研究, 对推动抗抑郁药物的研发具有重要意义。本实验从保山产赤芝中分离鉴定出2个新羊毛甾烷三萜(图1), 并从体内外研究评估了其抗抑郁活性。
结果
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1　结构鉴定
化合物1: 黄色固体。13C NMR、DEPT谱(表1)及HRESIMS确定其分子式为C30H40O5, 不饱和度为11。化合物1的1H NMR谱(表1)显示7个甲基(δH 0.96, 0.98, 1.09, 1.10, 1.21, 1.50, 1.82)和13C和DEPT谱(表1)显示30个碳信号, 包括7个甲基, 6个亚甲基, 6个次甲基(2个sp2杂化和1个含氧次甲基), 和11个非质子碳(包括2个酮基, 1个酯基, 4个sp2杂化, 4个sp3杂化)。分析化合物1的1D NMR数据, 发现其除了存在一个额外的酮基和亚甲基, 缺失了C-7位的亚甲基和C-16位的乙酰基, 以及在C-7、C-16和C-27周围化学位移发生局部变化外, 其余和(24Z)-16β-acetoxy-1,11α-epoxy-3-oxo-lanosta-1,8,24-trien-26-oic acid[10]非常相似。(A) C-7周围发生的化学位移变化: C-6 (δC 18.3 in CDCl3 → δC 36.9 in methanol-d4), C-7 (δC 25.8 in CDCl3 → δC 200.6 in methanol-d4)和C-9 (δC 132.2 in CDCl3 → δC 156.3 in methanol-d4)均向低场区方向移动。上述化学位移变化结合C-7 (δC 200.6)的化学位移推断C-7位存在一个酮基取代了原来的亚甲基。在1H-1H COSY谱(图2)中观察到H-5/H2-6的相关和HMBC谱(图2)中观察到H-5, H2-6/C-7的相关证实了推断A; (B) C-16周围发生的化学位移变化: C-15 (δC 39.3 in CDCl3 → δC 30.8 in methanol-d4), C-16 (δC 74.9 in CDCl3 → δC 29.2 in methanol-d4)和C-17 (δC 52.6 in CDCl3 → δC 49.5 in methanol-d4)均向高场区方向移动。上述化学位移变化结合C-16 (δC 29.2)的化学位移推断C-16位缺失了乙酰基, 由次甲基转变为亚甲基。在1H-1H COSY谱(图2)中观察到H2-15/H2-16/H-17的相关结合HMBC谱(图2)中观察到H3-21/C-17, C-20, C-22的相关证实了推断B; (C) C-27周围发生的化学位移变化: C-27 (δC 20.8 in CDCl3 → δC 12.4 in methanol-d4)和C-24 (δC 146.1 in CDCl3 → δC 144.1 in methanol-d4)向高场区移动, C-25 (δC 126.2 in CDCl3 → δC 128.9 in methanol-d4)向低场区移动推断是∆24(25)双键构型由Z转变为E引起。在ROESY谱(图2)中观察到H3-27/H2-23的相关而未观察到文献中H3-27/H-24的相关证实了推断C。此外, 在ROESY谱中观察到H-11/H3-18, H3-19和H3-18/H3-21, 表明H-11, H3-18, H3-19在同一侧, 定义为β构型, H-17为α构型。在ROESY谱中还观察到H3-30/H-5, H-17, 表明H-5, H-17和H3-30在同一平面, 为α构型, 因此, 确定了化合物1的相对构型为5R*, 10S*, 11R*, 13R*, 14R*, 17R*, 20R*。为了确定化合物1的绝对构型, 采用ECD计算的方法, 分别计算了(5R, 10S, 11R, 13R, 14R, 17R, 20R)-1和(5S, 10R, 11S, 13S, 14S, 17S, 20S)-1两种构型。结果表明, 化合物1的实测CD曲线与(5R, 10S, 11R, 13R, 14R, 17R, 20R)-1构型的ECD曲线拟合很好(图3)。因此, 确定了化合物1的绝对构型为5R, 10S, 11R, 13R, 14R, 17R, 20R, 命名为baoslingzhine X。
化合物2: 白色固体。13C NMR、DEPT谱(表1)及HRESIMS确定其分子式为C30H42O5, 不饱和度为10。化合物2的1H NMR谱(表1)包含7个甲基(δH 0.91, 0.98, 1.06, 1.09, 1.09, 1.14, 1.82)。13C和DEPT谱(表1)显示30个碳信号, 7个甲基, 7个亚甲基, 6个次甲基(1个sp2杂化和2个含氧次甲基), 和10个非质子碳(包括2个酮基, 1个酯基, 3个sp2杂化, 4个sp3杂化)。分析化合物2的1D NMR数据, 发现其除了存在一个额外的亚甲基和次甲基, 缺失了C-1位的烯季碳和C-2位的烯次甲基外, 其余均和化合物1非常相似。结合化合物的分子量推断∆1(2)双键还原为次甲基和亚甲基。在1H-1H COSY谱(图2)中观察到H-1/H2-2的相关, 结合H3-19/C-1 (δC 83.5), C-10和H2-2/C-3 (δC 207.5)的相关证实了上述结论, 因此, 确定了化合物2的平面结构。在ROESY谱(图2)中观察到H3-19/H-1, H-11和H3-18/H-20, H-11的相关推断H3-19, H-1, H-11和H3-18为β构型, H-17为α构型。此外, 在ROESY谱(图2)中观察到H3-30/H-5的相关推断H3-30和H-5为α构型。在ROESY谱(图2)中观察到H3-27/H2-23的相关推断∆24(25)双键为E构型。因此, 确定了化合物2的相对构型。为确定化合物2的绝对构型, 对2的两种构型(1S, 5R, 10S, 11R, 13R, 14R, 17R, 20R-2和1R, 5S, 10R, 11S, 13S, 14S, 17S, 20S-2)同样进行了ECD计算。结果表明, 化合物2的实测曲线与1S, 5R, 10S, 11R, 13R, 14R, 17R, 20R-2构型的ECD计算曲线拟合程度较高(图3)。因此, 确定了2的绝对构型为1S, 5R, 10S, 11R, 13R, 14R, 17R, 20R, 命名为baoslingzhine Y。
2　体内外抗抑郁活性评价
鉴于灵芝赤芝在抗神经炎症和抗抑郁症中具有强活性[11, 12], 本文对化合物1和化合物2分别进行体外抗炎、抗氧化及体内抗抑郁活性的测试。首先, 通过对一系列不同浓度的化合物1和2的细胞生存率的实验检测该类化合物的安全浓度。CCK-8的实验结果得到, 化合物1和2均在40 μmol·L-1内无毒。因此, 分别选择低浓度10 μmol·L-1和高浓度20 μmol·L-1进行后续细胞抗炎和抗氧化应激的检测。酶联免疫吸附实验结果发现, LPS可显著提高BV2细胞上清液中的促炎细胞因子IL-1β、IL-6和TNF-α的浓度, 而化合物1和2均以浓度依赖的方式降低BV2细胞因LPS引起的上述促炎因子的分泌(图4)。
活性氧荧光探针实验发现, LPS可显著提高BV2细胞中活性氧的荧光强度, 化合物1和2均以浓度依赖的方式降低BV2细胞因LPS引起的活性氧荧光强度的增加(图5A、B)。
一氧化氮试剂盒检测发现, LPS可显著提高BV2细胞中一氧化氮的水平, 10 μmol·L-1化合物1不改变一氧化氮的水平, 20 μmol·L-1化合物1则显著降低一氧化氮的水平, 而10和20 μmol·L-1化合物2均不改变一氧化氮的浓度(图5C)。以上实验结果表明, 化合物1和2均具有显著的抗神经炎症和抗氧化应激的活性。神经炎症作为抑郁症的核心症状之一, 许多抗抑郁药物均靶向神经炎症相关靶点[13]。
为了检测化合物1、2在抗抑郁中的作用, 本文使用经典的LPS刺激的神经炎症性抑郁症小鼠模型[14], 使用SSRI类传统抗抑郁药物氟西汀作为阳性对照, 设计体内实验。小鼠行为学数据表明, LPS模型、化合物1、2及氟西汀均不改变小鼠在爬竿实验和悬挂实验中的时间, 表明以上处理对小鼠的运动功能均无显著影响(图6)。基于上述研究结果, 本文检测一系列与抑郁症相关的指标, 包括体重变化、糖水实验、悬尾实验和强迫游泳。结果显示, LPS显著降低小鼠的体重和糖水偏好性, 并增加其在悬尾实验和强迫游泳中的不动时间。然而, 化合物1、2和氟西汀均显著性改善上述指标(图7)。以上结果表明, 化合物1、2具有显著的抗抑郁活性。
实验部分
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Anton Paar MCP-100 digital polarimeter型全自动数字旋光仪(Anton Paar公司); 圆二色谱仪(应用光物理公司); SP 5030型半制备高效液相色谱仪(北京赛谱锐思科技有限公司); AB SCIEX triple TOF X500R高分辨质谱仪(AB SCIEX公司); Bruker AV-500 MHz及AV-600 MHz核磁共振波谱仪(以TMS为内标, 德国Bruker公司); MCI gel CHP 20P填料(70~150 μm, 日本三菱公司); YMC gel ODS-A-HG填料(12 nm S-50 μm, 日本YMC公司); 半制备SEP Basic C18色谱柱(120A 5 μm, 250 mm × 10 mm, 北京赛谱锐思科技有限公司)。细胞培养箱(美国Thermo Fisher Scientific公司); 酶标仪(美国Bio-Tek公司); 超高分辨激光共聚焦显微LSM880 (德国Carl Zeiss Jena公司); 行为学仪器(上海欣软信息科技有限公司); VisuTrack行为学分析软件(上海欣软信息科技有限公司)。
灵芝药材于2018年4月购自云南省保山市滇滩镇胜利村, 由中国科学院昆明植物研究所杨祝良教授鉴定为赤芝的干燥子实体, 样品标本(CHYX0619)已存放在广东省深圳市深圳大学药学院。
提取分离    取赤芝干燥子实体500.0 kg, 粉碎后用80%乙醇回流提取(2 × 3 000 L × 3 h), 合并回流液, 减压浓缩至无乙醇味, 得粗浸膏(约24.6 kg)。将总浸膏用温水混悬后, 加等体积乙酸乙酯萃取4次, 减压浓缩得乙酸乙酯部位浸膏(11.5 kg)。其经MCI gel CHP 20P反相色谱柱以甲醇-水系统(40%~100%)梯度洗脱, TLC监测合并相同部分, 得到17个组分Fr. A~Q。经HPLC-DAD-ESI/MS检测分析, 选择Fr. K进行分离纯化。
Fr. K (379.0 g)经MCI gel CHP 20P柱梯度洗脱(甲醇-水, 30%~100%), 得到7个组分(Fr.K.1~Fr.K.7)。Fr.K.7 (36.2 g)经凝胶洗脱后, 得到3个亚组分(Fr.K.7.1~Fr.K.7.3), 其中第2个亚组分Fr.K.7.2 (21.0 g)经RP-18柱(甲醇-水, 70%~100%)分离得到6段(Fr.K.7.2.1~Fr.K.7.2.6)。Fr.K.7.2.5 (7.1 g)经凝胶分离后又得到3个组分(Fr.K.7.2.5.1~Fr.K.7.2.5.3), 其中Fr.K.7.2.5.3 (855.0 mg)经凝胶再次分离后又得到2个组分(Fr.K.7.2.5.3.1和Fr.K.7.2.5.3.2)。Fr.K.7.2.5.3.1 (387.8 mg)经半制备液相HPLC反复纯化得到化合物1 (甲醇-水, 81%, 水中含0.05% TFA, 流速: 3 mL·min-1, tR = 20.1 min, 1.60 mg)和化合物2 (甲醇-水, 79%, 水中含0.05% TFA, 流速: 3 mL·min-1, tR = 17.9 min, 2.73 mg)。
结构鉴定    化合物1, 白色固体, 溶于甲醇。[α]${}_{\mathrm{D}}^{20} $ +145.4 (c 0.04, MeOH); UV (MeOH) λmax (log ε): 256 (4.29), 218 (4.29) nm; CD (MeOH) λmax (Δε): 309 (-14.6), 259 (+59.6) nm; HRESIMS m/z 481.293 4 [M+H]+ (Calcd. for C30H41O5, 481.294 9); 1H and 13C NMR数据见表1。
化合物2, 白色固体, 溶于甲醇。[α]${}_{\mathrm{D}}^{20} $ +96.5 (c 0.06, MeOH); UV (MeOH) λmax (log ε): 218 (4.58) nm; CD (MeOH) λmax (Δε): 295 (+10.0), 248 (-12.0), 222 (+15.0), 218 (+13.4), 212 (+18.4) nm; HRESIMS m/z 483.309 7 [M+H]+ (Calcd. for C40H43O5, 483.310 5); 1H和13C NMR数据见表1。
细胞培养    BV2小鼠小胶质细胞培养于含10%胎牛血清(FBS, Gibco, 货号2094468CP)的DMEM/F-12培养基(Thermo Fisher Scientific, 货号A4192001)中, 置于5% CO2的湿润培养箱内。
细胞活力检测    将BV2细胞均匀接种于96孔板内, 贴壁过夜后, 用不同浓度的化合物处理细胞, 培养24 h后, 按照CCK-8试剂盒(翌圣生物, 货号40203ES60)的实验方法检测并计算细胞存活率。
酶联免疫吸附实验、一氧化氮(NO)检测和细胞内ROS含量测定    将BV2细胞接种于24孔板, 过夜贴壁后, 分别用不同浓度的化合物处理14 h后, 与1 μg·mL-1脂多糖(LPS, Solarbio, L8880)共同孵育10 h, 收集细胞培养液, 4 ℃条件下离心15 min (3 000 r·min-1), 取上清进行后续实验。上清液按ELISA试剂盒(酶联生物)说明书和一氧化氮试剂盒(Beyotime, S0021S)说明书进行检测。
随后加入500 μL 10 μmol·L-1的DCFH-DA (Yeasen, 货号50101ES01), 在37 ℃细胞培养箱内避光孵育30 min, 于显微镜下观察ROS荧光强度。
动物及分组    本研究经深圳大学动物伦理与福利委员会批准(批准号: IACUC-202400148)。实验采用8周龄C57BL/6雄性小鼠, 购自广东至远生物医药科技有限公司许可证[SCXK (粤) 2023-0066], 购买后于塑料饲养笼中适应饲养1周。饲养环境温度维持在24~26 ℃、湿度60%。研究共包括两部分实验。实验分为五组。正常对照组(NC): C57小鼠腹腔注射生理盐水(100 μL每10 g体重)。LPS模型组: C57小鼠腹腔注射脂多糖(LPS, 1 mg·kg-1), 连续给药2天。LPS+化合物1/2组: 在第二次LPS注射4 h后, 通过鼻滴给药化合物1或2 (浓度2 mg·kg-1)。LPS+氟西汀组: 在LPS给药后4 h, 每日腹腔注射氟西汀(阳性药, 10 mg·kg-1)。
行为学检测    各组小鼠分别在相应处理后1 h依次进行爬竿实验/悬挂实验、蔗糖偏好实验、悬尾实验和强迫游泳实验, 行为学实验流程参考先前已发表的文献[15]。爬竿测试: 将小鼠放置在直径为10 mm、高度为50 cm的杆子顶端, 记录其从顶部移动到底部所需时间。悬挂测试: 将小鼠置于网格中央并反向观察其掉落时间(最大时间限制为90 s, 超过90 s则为统计值90 s)。悬尾实验(tail suspension test, TST): 将小鼠放置于测试实验室1 h以适应环境后, 用胶带缠住尾巴并固定在离地面约40 cm的不透明长方形塑料隔间(长20 cm、宽11.5 cm、高55 cm), 记录5 min内小鼠的摇摆时间, 使用VisuTrack Video软件进行分析。强迫游泳实验(forced swimming test, FST): 实验开始前准备高50 cm、直径20 cm的透明圆柱形容器, 盛有约25 cm高度的水, 水温约为25 ℃。小鼠适应环境1 h后, 将其放入水中5 min, 通过VisuTrack Video软件(上海欣软)分析小鼠漂浮不动的时间。糖水偏好实验(sucrose preference test, SPT): 将小鼠禁水过夜后, 单笼饲养, 同时给予一瓶水和一瓶浓度为1%的蔗糖水, 10 h后取下, 分别记录水和1%蔗糖水的消耗量。小鼠的糖水偏好性= 1%蔗糖水消耗量/ (水消耗量+ 1%蔗糖水消耗量) × 100%。
统计学分析    实验数据通过GraphPad Prism 10.4软件进行处理, 以平均值± SEM (means ± SEM)表示。采用单因素方差分析(one-way ANOVA检验)方法进行各组间差异对比分析。P < 0.05则组间差异具有统计学意义。

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doi: 10.16438/j.0513-4870.2026-0112
  • 接收时间：2026-01-27
  • 首发时间：2026-09-21
  • 出版时间：2026-07-12
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  • 收稿日期：2026-01-27
  • 修回日期：2026-04-05
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    1云南中医药大学, 基础医学院, 云南 昆明 650500
    2深圳大学医学部药学院, 中医药守正创新研究院, 广东 深圳 518060

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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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