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Article(id=1308455899765428708, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308455335962894804, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2025-0009, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1735920000000, receivedDateStr=2025-01-04, revisedDate=1739980800000, revisedDateStr=2025-02-20, acceptedDate=null, acceptedDateStr=null, onlineDate=1789889796439, onlineDateStr=2026-09-20, pubDate=1749657600000, pubDateStr=2025-06-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789889796439, onlineIssueDateStr=2026-09-20, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789889796439, creator=13701087609, updateTime=1789889796439, updator=13701087609, issue=Issue{id=1308455335962894804, tenantId=1146029695717560320, journalId=1189982191388893191, year='2025', volume='60', issue='6', pageStart='1573', pageEnd='1964', issueExtLink='null', onlineDate='null', pubDate='1749657600000', pubDateStr='2025-06-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789889662017, creator='13701087609', updateTime=1789893590327, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1308471812564734358, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308455335962894804, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1308471812564734359, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308455335962894804, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1843, endPage=1846, ext={EN=ArticleExt(id=1308455899983532517, articleId=1308455899765428708, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=A new pregnane-type steroid from Munronia unifoliolate Oliv., columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

Using separation methods such as silica gel, MCI, ODS, Sephadex LH-20 gel column chromatography, and high-performance liquid chromatography, two C21 steroid components were isolated and identified from the traditional medicinal plant Munronia unifoliolata Oliv. (commonly known as "Aituotuo"), which is used for treating rheumatic joint pain and traumatic injuries. The structures of these compounds were identified as 2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) and 2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2) using comprehensive structural identification methods, including UV, IR, high-resolution mass spectrometry, and one-dimensional and two-dimensional nuclear magnetic resonance. Compounds 1 and 2 were reported for the first time as pregnane-type components from M. unifoliolata, with compound 1 being a new compound. The results of the bioactivity screening indicated that both compound 1 and compound 2 did not inhibit nitric oxide (NO) production and showed no activity in reversing multidrug resistance.

, authors=Meng-meng YU1, Yun-peng SUN2, Jun LUO1, *, Ling-yi KONG1, *, authorsList=Meng-meng YU, Yun-peng SUN, Jun LUO, Ling-yi KONG, authorCompany=null, correspAuthors=Jun LUO, Ling-yi KONG, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2025 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=1308455900776255978, articleId=1308455899765428708, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=单叶地黄连中一个新的孕甾烷成分, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

采用硅胶、MCI、ODS、Sephadex LH-20凝胶柱层析、高效制备液相色谱等分离手段, 从用于风湿关节痹痛和跌打损伤治疗的民间药物“矮坨坨”的基源植物单叶地黄连(Munronia unifoliolata Oliv.) 中分离并鉴定了两个C21甾体类成分。综合运用紫外、红外、高分辨质谱、一维和二维核磁共振等结构鉴定方法, 鉴定其结构分别为2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) 和2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2)。化合物1和2为首次从单叶地黄连中报道的孕甾烷类成分, 且化合物1为新化合物。生物活性筛选结果显示, 化合物1和2均不抑制NO生成, 且无逆转多药耐药活性。

, authors=于萌萌1, 孙云鹏2, 罗俊1, *, 孔令义1, *, authorsList=于萌萌, 孙云鹏, 罗俊, 孔令义, authorCompany=null, correspAuthors=罗俊, 孔令义, authorNote=

作者贡献

于萌萌负责化合物的提取分离、结构鉴定、数据处理和论文撰写; 孙云鹏参与结构鉴定和数据分析处理部分工作; 孔令义、罗俊对课题研究提供指导和提出写作的建议。

, correspAuthorsNote=
罗俊, E-mail:
孔令义, Tel: 86-25-83271405, E-mail:
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No.12
δH (J, Hz)δCδH (J, Hz)δC
11.66, m41.11.72, m40.7
23.75, d (11.4)69.23.78, d (11.2)69.2
33.96, s69.43.98, s69.4
41.59, m34.31.60, m34.3
1.53, m
51.54, m38.41.55, m38.4
61.34, m27.61.34, m27.6
1.22, m
71.91, m31.41.90, m31.4
81.70, m30.830.8
90.86, m54.80.87, m54.8
1037.237.2
111.47, m20.91.51, m20.9
1.21, m1.23, m
121.84, m40.941.1
1342.342.2
140.97, m60.60.94, m60.5
154.30. t (5.8)70.74.28, t (5.5)70.7
162.33, m38.62.28, m38.5
1.41, m
171.73, m55.61.62, m55.6
180.93, s15.40.92, s15.4
190.79, s12.50.83, s12.2
205.24, dd (6.0, 10.4)73.15.05, dd (6.0, 10.3)72.3
211.28, d (6.1)20.11.18, d (6.0)20.1
1′165.8167.4
2′130.9129.2
3′/7′8.04, d (7.4)129.86.84, d (6.9)137.2
4′/6′7.44, t (7.7)128.51.80, d (7.3)14.5
5′7.55, t (7.4)132.91.83, s12.5
), ArticleFig(id=1308497593659916937, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308455899765428708, language=CN, label=Tab.1, caption=

1H (600 MHz, J in Hz) and 13C (150 MHz in CDCl3) NMR data of compounds 1 and 2

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No.12
δH (J, Hz)δCδH (J, Hz)δC
11.66, m41.11.72, m40.7
23.75, d (11.4)69.23.78, d (11.2)69.2
33.96, s69.43.98, s69.4
41.59, m34.31.60, m34.3
1.53, m
51.54, m38.41.55, m38.4
61.34, m27.61.34, m27.6
1.22, m
71.91, m31.41.90, m31.4
81.70, m30.830.8
90.86, m54.80.87, m54.8
1037.237.2
111.47, m20.91.51, m20.9
1.21, m1.23, m
121.84, m40.941.1
1342.342.2
140.97, m60.60.94, m60.5
154.30. t (5.8)70.74.28, t (5.5)70.7
162.33, m38.62.28, m38.5
1.41, m
171.73, m55.61.62, m55.6
180.93, s15.40.92, s15.4
190.79, s12.50.83, s12.2
205.24, dd (6.0, 10.4)73.15.05, dd (6.0, 10.3)72.3
211.28, d (6.1)20.11.18, d (6.0)20.1
1′165.8167.4
2′130.9129.2
3′/7′8.04, d (7.4)129.86.84, d (6.9)137.2
4′/6′7.44, t (7.7)128.51.80, d (7.3)14.5
5′7.55, t (7.4)132.91.83, s12.5
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单叶地黄连中一个新的孕甾烷成分
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于萌萌 1 , 孙云鹏 2 , 罗俊 1, * , 孔令义 1, *
药学学报 | 研究论文 2025,60(6): 1843-1846
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药学学报 |研究论文 2025 , 60 (6) : 1843 -1846
单叶地黄连中一个新的孕甾烷成分
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于萌萌1, 孙云鹏2, 罗俊1, * , 孔令义1, *
作者信息
  • 1中国药科大学中药学院, 江苏 南京 210000
  • 2安徽中医药大学药学院, 安徽 合肥 230000
通讯作者:
罗俊, E-mail:
孔令义, Tel: 86-25-83271405, E-mail:
作者简介:

作者贡献

于萌萌负责化合物的提取分离、结构鉴定、数据处理和论文撰写; 孙云鹏参与结构鉴定和数据分析处理部分工作; 孔令义、罗俊对课题研究提供指导和提出写作的建议。

A new pregnane-type steroid from Munronia unifoliolate Oliv.
Meng-meng YU1, Yun-peng SUN2, Jun LUO1, * , Ling-yi KONG1, *
Affiliations
  • 1School of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing 210000, China
  • 2School of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230000, China
出版时间: 2025-06-12 doi: 10.16438/j.0513-4870.2025-0009
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摘要
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采用硅胶、MCI、ODS、Sephadex LH-20凝胶柱层析、高效制备液相色谱等分离手段, 从用于风湿关节痹痛和跌打损伤治疗的民间药物“矮坨坨”的基源植物单叶地黄连(Munronia unifoliolata Oliv.) 中分离并鉴定了两个C21甾体类成分。综合运用紫外、红外、高分辨质谱、一维和二维核磁共振等结构鉴定方法, 鉴定其结构分别为2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) 和2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2)。化合物1和2为首次从单叶地黄连中报道的孕甾烷类成分, 且化合物1为新化合物。生物活性筛选结果显示, 化合物1和2均不抑制NO生成, 且无逆转多药耐药活性。

单叶地黄连  /  矮坨坨  /  提取分离  /  孕甾烷

Using separation methods such as silica gel, MCI, ODS, Sephadex LH-20 gel column chromatography, and high-performance liquid chromatography, two C21 steroid components were isolated and identified from the traditional medicinal plant Munronia unifoliolata Oliv. (commonly known as "Aituotuo"), which is used for treating rheumatic joint pain and traumatic injuries. The structures of these compounds were identified as 2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) and 2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2) using comprehensive structural identification methods, including UV, IR, high-resolution mass spectrometry, and one-dimensional and two-dimensional nuclear magnetic resonance. Compounds 1 and 2 were reported for the first time as pregnane-type components from M. unifoliolata, with compound 1 being a new compound. The results of the bioactivity screening indicated that both compound 1 and compound 2 did not inhibit nitric oxide (NO) production and showed no activity in reversing multidrug resistance.

Munronia unifoliolata  /  Aituotuo  /  extraction and isolation  /  pregnane
于萌萌, 孙云鹏, 罗俊, 孔令义. 单叶地黄连中一个新的孕甾烷成分. 药学学报, 2025 , 60 (6) : 1843 -1846 . DOI: 10.16438/j.0513-4870.2025-0009
Meng-meng YU, Yun-peng SUN, Jun LUO, Ling-yi KONG. A new pregnane-type steroid from Munronia unifoliolate Oliv.[J]. Acta Pharmaceutica Sinica, 2025 , 60 (6) : 1843 -1846 . DOI: 10.16438/j.0513-4870.2025-0009
正文
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地黄连属(Munronia) 是楝科植物中少见的矮小灌木或半灌木, 全世界15种, 我国产8种2变种, 主要分布于我国中部、西南部和南部地区[1]。多种地黄连属植物以“矮坨坨”之名在民间广泛运用, 具有清热解毒、活血止痛、祛风通络、截疟等功效[2]。单叶地黄连(Munronia unifoliolata Oliv.), 主要生长在我国的西南地区, 全株入药, 味微苦、涩、凉, 主治跌打损伤、风湿关节痛、痈肿疔疮。目前有关地黄连属植物的研究较少, 且大部分集中于云南地黄连(M. delavayi Franch) 和矮陀陀(M. henryi Harms), 从中分离得到了大量柠檬苦素成分[3-7]。前期, 课题组对来自多种楝科药用植物中的柠檬苦素类成分开展了较为系统的研究[8-12], 其中从单叶地黄连中分离得到个39个新柠檬苦素及前体三萜类化合物[13-15], 并发现了柠檬苦素类化合物具有良好的抑制NLRP3炎症小体激活的抗炎活性[13, 16]。为了进一步全面阐明单叶地黄连中的化学成分类型, 作者对前期的剩余浸膏进行了再研究。从二氯甲烷部位又分离了2个少见的孕甾烷类C21甾体成分(图1), 并鉴定为2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane (1) 和2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane (2)。化合物1为新化合物[17], 且1和2为首次从单叶地黄连中报道孕甾烷类成分。生物活性筛选结果显示, 化合物1和2均不抑制NO生成, 且无逆转多药耐药活性。
结果与讨论
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1　化合物结构鉴定
化合物1为白色无定形粉末。HR-ESI-MS (实测值: m/z 479.276 9 [M+Na]+, 计算值: 479.276 8) 提示其分子式为C28H40O5, 不饱和度为9。1H NMR谱(表1) 显示一组单取代苯的信号[δH 8.04 (2H, d, J = 7.4 Hz), 7.55 (2H, t, J = 7.4 Hz), 7.44 (1H, t, J = 7.7 Hz)], 4个连氧次甲基信号[δH 5.24 (1H, dd, J = 6.0, 10.4 Hz), 4.30 (1H, t, J = 5.8 Hz), 3.96 (1H, s), 3.75 (1H, d, J = 11.4 Hz)], 1个双峰甲基[δH 1.28 (3H, d, J = 6.1 Hz)] 和2个单峰甲基[δH 0.93 (3H, s), 0.79 (3H, s)]。13C NMR谱显示28个碳信号(2对苯环重叠信号), 其中包括1个酯羰基信号(δC 165.8), 6个苯环碳信号(δC 132.9, 130.9, 129.8, 129.8, 128.5, 128.5) 以及4个连氧碳信号(δC 73.1, 70.7, 69.4, 69.2)。HMBC谱(图2) 中H-3′与C-1′的相关信号证明苯甲酰基的存在。除去苯甲酰基的碳信号, 1结构中还有21个碳, 结合以上数据分析, 确定1为孕甾烷类化合物, 并且与2非常相似[18]。对比两者的核磁谱图, 发现它们之间仅取代基的不同。H-20 (δH 5.24, dd, J = 6.0, 10.4 Hz) 与C-1′ (δC 165.8) 的HMBC相关信号表明苯甲酰基取代在C-20位。1的相对构型通过孕甾烷的骨架构型和ROESY谱确定。在ROESY谱中, H-2 (δH 3.78, d, J = 11.2 Hz)/Me-19 (δH 0.79, s) 的相关表明H-2/Me-19的取代是β取向, 此外H-2 (δH 3.75, d, J = 11.4 Hz)/H-3 (δH 3.96, s) 存在相关, 且其与化合物2中H-2 (δH 3.78, d, J = 11.2 Hz), H-3 (δH 3.98, s) 的核磁信号近似, 以上数据表明其H-2, H-3均为β构型。同时, Me-21 (δH 1.28, d, J = 6.1 Hz)/H-17 (δH 1.73, m), Me-21/H-16α (δH 2.33, m) 的相关信号说明Me-21/H-17/H-16α为α型。H-20 (δH 5.24, dd, J = 10.4, 6.0 Hz)/H-16β (δH 1.41, m), H-20/Me-18 (δH 0.93, s) 的相关信号说明H-20/H-16β/Me-18为β构型。H-14 (δH 0.97, m)/H-15 (δH 4.30, t, J = 5.8 Hz)/H-16α的相关信号说明OH-15是β取向的。通过仔细对比文献[18-20]的光谱数据和H-17, H-18, H-20和H-21四个关键氢的ROESY相关峰, 以及ECD吸收曲线和旋光值, 化合物1的绝对构型与文献报道的2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane一致, 特别是C-20被定为S构型。综上所述, 1的结构确定如上图所示, 并命名为2α, 3α, 15β-trihydroxy-20(S)-benzoyl-pregnane, 经文献检索为一新化合物。
化合物2为白色无定形粉末。1H NMR谱(表1) 显示一组惕铬酰基信号[6.84 (1H, q, J = 6.9 Hz), 1.84 (3H, s), 1.80 (3H, d, J = 7.3 Hz)], 4个连氧次甲基信号[5.05 (1H, dd, J = 6.0, 10.3 Hz), 4.28 (1H, t, J = 5.5 Hz), 3.98 (1H, s), 3.78 (1H, d, J = 11.2 Hz)], 1个二重峰甲基信号和2个单峰甲基信号, 以上提示2为孕甾烷类化合物。通过与文献[18]数据对比, 确定化合物2为2α, 3α, 15β-trihydroxy-20(S)-tigloyl-pregnane。
2　活性评价
单叶地黄连作为传统中药, 主治跌打损伤、风湿关节痛、痈肿疔疮, 具有抗炎的功效。本文对化合物1和2抑制脂多糖(LPS) 诱导的小鼠单核巨噬细胞(RAW264.7) 释放一氧化氮(NO) 的能力进行了测试, 初步评估其抗炎活性。结果表明, 化合物1和2表现出较弱的NO抑制活性(IC50 > 40 μmol·L-1)。同时, 本文采用MTT法测试化合物1和2对MCF/7和MCF-7/DOX细胞的细胞毒性, 结果显示上述化合物对耐药细胞株均无明显细胞毒性(IC50 > 30 μmol·L-1)。因此, 进一步测试了上述化合物对MCF-7/DOX细胞株多药耐药的逆转作用。结果表明, 化合物1和2也不表现逆转多药耐药活性, 逆转倍数小于1.0。
实验部分
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P-1020型自动旋光测定仪, J-810型圆二色谱仪(日本JASCO公司); LC-6AD及LC-20AR型HPLC制备液相色谱仪, 配备SPD-20A双波长检测器, 色谱柱为Shim-pack RP-C18, 5/10 μm, 20 mm × 200 mm, UV-2450型UV/Vis分光光度计(日本SHIMADZU公司); 1100 Series LC/MSD Trap型低分辨质谱仪、UPLC-Q-TOF 6520B型高分辨质谱仪、1260型HPLC分析液相色谱仪、ZORBAX Eclipse XDB-C18型色谱柱(美国Agilent公司); Tensor-27型KBr压片红外光谱仪、Avance Ⅲ-500和AV-600核磁共振波谱仪(TMS为内标, 德国Bruker公司)。
100~200和200~300目柱层析硅胶和硅胶薄层板(GF254) (青岛海洋化工有限公司); ODS: 50 µm (日本YMC株式会社); 40~70 µm Sephadex LH-20凝胶(瑞典Amersham Pharmacia Biotech AB公司); MCI树脂(日本三菱化学株式会社); 试剂: 甲醇、乙腈、二氯甲烷、石油醚、乙酸乙酯等(江苏汉邦科技有限公司)。
野生单叶地黄连干燥全草(12.8 kg), 2019年8月采自云南省丽江市。由中国药科大学生药学教研室张勉教授鉴定为楝科地黄连属植物单叶地黄连(Munronia unifoliolata Oliv.), 凭证标本(20190801) 存放于中国药科大学天然药物化学教研室。
1　提取分离
单叶地黄连干燥全草(12.8 kg), 粉碎, 95%乙醇回流提取4次, 首次3 h, 之后每次2 h, 提取液滤过后减压浓缩至无醇味, 得总提浸膏(2.4 kg)。粗浸膏用水混悬后, 二氯甲烷萃取3次, 萃取液减压浓缩得二氯甲烷部位。二氯甲烷部位加50%甲醇-水混悬, 用石油醚反萃3次, 得石油醚部位(350 g) 和反萃后的二氯甲烷部位(260 g)。二氯甲烷部位(260 g) 经硅胶柱色谱, 二氯甲烷-甲醇系统梯度洗脱(100∶1~0∶100), 得7个组分。其中20∶1洗脱组分(Fr.E, 13.5 g) 经MCI柱色谱, 甲醇-水系统梯度洗脱(60∶40~100∶0), 洗脱液经薄层检识合并相同组分, 得到5个组分(Fr.E1~Fr.E5)。Fr.E3组分(4.5 g) 经中压柱色谱(ODS), 甲醇-水系统梯度(40∶60~100∶0) 洗脱, 合并相同组分后得到8个亚组分(Fr.E3a~Fr.E3h), Fr.E3e (236 mg) 经Sephadex LH-20凝胶(甲醇) 纯化后, 再经高效制备液相色谱(甲醇-水, 70∶30) 制备得化合物1 (7 mg, 40.5 min)。Fr.E3f (189 mg) 经高效制备液相色谱(乙腈-水, 60∶40) 制备得化合物2 (17 mg, 32.6 min)。
2　结构鉴定
2α, 3α, 15β-Trihydroxy-20(S)-benzoyl-pregnane (1) 白色无定形粉末; [α]$ {}_{\mathrm{D}}^{19} $ -23.3 (c 0.10, MeOH); UV (MeOH) λmax (log ε) 203 (3.87) nm, 227 (3.88) nm; ECD (MeOH) λmax (Δε) 230 (+4.94) nm; IR (KBr) νmax 3 436, 2 930, 1 713, 1 451, 1 383, 1 278, 1 121, 1 027 cm-1; 1H and 13C NMR数据见表1; HRESIMS m/z 479.276 9 [M+Na]+ (calcd. for C28H40O5Na, 479.276 8)。
2α, 3α, 15β-Trihydroxy-20(S)-tigloyl-pregnane (2) 白色无定形粉末; [α]$ {}_{\mathrm{D}}^{19} $ -8.5 (c 0.10, MeOH); ESI-MS 557.3 [M+Na]+; 分子式: C26H42O5; 1H and 13C NMR数据见表1。
3　活性评价
将取对数生长期状态良好的RAW264.7细胞接种到96孔板中, 细胞贴壁后用3.125、6.25、12.5、25、50 μmol·L-1浓度的化合物孵育细胞, 每组设置三个复孔。孵育一定时间后, 每孔加入10 μL的MTT试剂并孵育4 h。然后吸去上清液, 并通过持续摇动加入DMSO (150 μL) 10 min。检测570 nm波长下的吸光度值(A)。计算细胞抑制率并表示为IC50值[抑制率= (A空白 - A样品) / A空白]。
取对数生长期状态良好的RAW264.7细胞接种到96孔板上(每孔100 μL), 在37 ℃、5% CO2的恒温培养箱中培养18 h。不加或者加入40 μmol·L-1化合物或40 μmol·L-1阳性药L-单甲基精氨酸孵育细胞, 每组设置三个复孔, 给药1 h后加入LPS刺激NO生成, 继续培养18 h。每孔取50 μL上清液, 按NO试剂盒说明书流程操作, 用酶标仪检测540 nm波长下各孔的A值。计算NO抑制率并表示为IC50值[抑制率= (A模型 - A样品) /(A模型 - A空白)]。
取对数生长期状态良好的人乳腺癌细胞(MCF-7) 和多柔比星耐药的人乳腺癌细胞(MCF-7/DOX), 接种在96孔板上于恒温培养箱中培养24 h。加入不同浓度的多柔比星后, 不加或者加入30或10 μmol·L-1的化合物或阳性药维拉帕米。培养48 h后, 向每孔加入20 μL MTT并孵育4 h。然后吸去上清液, 并通过持续摇动加入DMSO (150 μL) 10 min。检测570 nm波长下的A值。耐药指数= IC50 (MCF-7/DOX cell) / IC50 (MCF-7 cell)。逆转倍数= IC50 (MCF-7/DOX cell空白) / IC50 (MCF-7 cell给药)。

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doi: 10.16438/j.0513-4870.2025-0009
  • 接收时间：2025-01-04
  • 首发时间：2026-09-20
  • 出版时间：2025-06-12
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  • 收稿日期：2025-01-04
  • 修回日期：2025-02-20
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    1中国药科大学中药学院, 江苏 南京 210000
    2安徽中医药大学药学院, 安徽 合肥 230000

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2种不同金属材料的力学参数
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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
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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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