民族药阴地蕨抗结核活性筛选及化学成分分析

doi:10.19982/j.issn.1000-6621.20260021

中国防痨杂志 ›› 2026, Vol. 48 ›› Issue (6): 787-794.doi: 10.19982/j.issn.1000-6621.20260021

民族药阴地蕨抗结核活性筛选及化学成分分析

李静¹, 陈双双²^,³, 陈春华¹, 杨国祥¹, 杨承闹¹, 郑禄云¹, 谭承建¹, 李传友²^,³, 李洁²^,³, 周培富¹(), 代小伟²^,³()

¹ 贵州民族大学民族医药学院, 中华人民共和国国家民族事务委员会贵州少数民族医药资源开发与利用重点实验室, 贵阳 550025
² 北京市疾病预防控制中心结核病实验室, 北京 100035
³ 首都医科大学公共卫生学院, 北京 100069

收稿日期:2026-01-12 出版日期:2026-06-10 发布日期:2026-05-25
通信作者: 周培富,代小伟 E-mail:zhoupeifu@sina.com;happydaixw@126.com
基金资助:
国家自然科学基金项目(82260404);贵州省基础研究计划项目(自然科学)面上项目(黔科合基础MS〔2025〕221)

Screening of anti-tuberculosis activity and analysis of chemical constituents of the Ethnomedicine Botrychium ternatum

Li Jing¹, Chen Shuangshuang²^,³, Chen Chunhua¹, Yang Guoxiang¹, Yang Chengnao¹, Zheng Luyun¹, Tan Chengjian¹, Li Chuanyou²^,³, Li Jie²^,³, Zhou Peifu¹(), Dai Xiaowei²^,³()

¹ Key Laboratory of the Development and Utilization of Guizhou Minority Medical Resource, National Ethnic Affairs Commission, School of Chinese Ethnic Medicine, Guizhou Minzu University, Guiyang 550025, China
² Tuberculosis Laboratory, Beijing Center for Disease prevention and control, Beijing 100035, China
³ China School of Public Health, Capital Medical University, Beijing 100069, China

Received:2026-01-12 Online:2026-06-10 Published:2026-05-25
Contact: Zhou Peifu,Dai Xiaowei E-mail:zhoupeifu@sina.com;happydaixw@126.com
Supported by:
National Natural Science Foundation of China(82260404);General Program of Natural Science, Basic Research Program of Guizhou Province (Qian Ke He Jichu-MS 〔2025〕221)

摘要/Abstract

摘要：

目的: 筛选具有抗结核活性的民族药物,评估其活性并解析化学成分,为结核病治疗提供新思路。方法: 通过文献研究筛选15种民族药,制备总浸膏,采用固体琼脂纸片扩散法初筛抗耻垢分枝杆菌(Mycobacterium smegmatis, M.sm)活性;选择活性最佳药物提取有机相分离物(石油醚、二氯甲烷、正丁醇),通过刃天青微量肉汤稀释法检测对M.sm和卡介苗(Bacillus Calmette-Guérin,BCG)的最低抑菌浓度(minimum inhibitory concentration,MIC),并用微量肉汤稀释法检测对结核分枝杆菌标准株H37Rv及临床分离株(包括耐药菌株和敏感菌株)的活性;通过高效液相色谱-质谱(high-performance liquid chromatography-mass spectrometry,HPLC-MS)分析阴地蕨总浸膏及石油醚相分离物化学成分,初步分析其潜在抗结核活性机制。结果: 查询整理的15种民族药中,10种对M.sm有抑菌活性,阴地蕨活性最强[抑菌圈直径为(18.33±4.04)mm]。刃天青法药物敏感性试验检测阴地蕨总浸膏及其3个有机相分离物对M.sm和BCG的MIC显示,阴地蕨总浸膏和石油醚相分离物抑菌效果均明显,两者对M.sm的MIC均为1875μg/ml、对BCG的MIC分别为2μg/ml和1μg/ml;微量肉汤稀释法药物敏感性试验检测阴地蕨总浸膏和石油醚相分离物对11株结核分枝杆菌临床分离株MIC主要集中在14~58μg/ml,其中石油醚相分离物MIC最低达到14μg/ml。HPLC-MS分析阴地蕨总浸膏鉴别出44种化合物,石油醚相分离物鉴别出33种化合物。结论: 阴地蕨在15种民族药中抗M.sm活性最佳,进一步活性评价表明,阴地蕨特别是其石油醚相分离物具有显著抗结核活性。研究不仅验证了从民族药中筛选抗结核药物的可行性,其成分分析结果为开发新型抗结核药物提供了重要的候选物质与科学依据。

关键词: 分枝杆菌,结核, 药物活性筛查, 最低抑菌浓度, 民族药, 成分分析, 阴地蕨

Abstract:

Objective: To screen ethnomedicines with anti-tuberculosis activity, evaluate their efficacy and chemical composition, and provide new insights for tuberculosis treatment. Methods: Fifteen ethnomedicines were selected through literature review, and their total extracts were prepared. Initial screening for activity against Mycobacterium smegmatis (M.sm) was performed using the solid agar disk diffusion method. The medicine with the best activity was further extracted to obtain organic solvent fractions (petroleum ether, dichloromethane, n-butanol). The minimum inhibitory concentrations (MIC) of these fractions against M.sm and Bacillus Calmette-Guérin (BCG) were determined by the resazurin microtiter broth dilution assay. The activity against the standard strain H37Rv and clinical isolates (including drug-resistant and drug-sensitive strains) of Mycobacterium tuberculosis was evaluated via the microbroth dilution method. The chemical constituents of the total extract and petroleum ether fraction from Botrychium ternatum were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS) to preliminarily explore the potential anti-tuberculosis components. Results: Among the 15 ethnomedicines screened, 10 exhibited inhibitory activity against M.sm, with Botrychium ternatum showing the strongest activity (inhibition zone diameter: 18.33±4.04 mm). Resazurin assay results indicated significant inhibitory effects for the total extract and the petroleum ether fraction of Botrychium ternatum against M.sm and BCG, with MICs of 1875 μg/ml against M.sm for both, and MIC of 2 μg/ml, and 1 μg/ml against BCG for total extract and the petroleum ether fraction, respectively. Microbroth dilution testing against 11 clinical isolates of Mycobacterium tuberculosis showed that the MICs for the total extract and the petroleum ether fraction mainly ranged from 14 to 58 μg/ml, with the lowest MIC for the petroleum ether fraction reaching 14 μg/ml. HPLC-MS analysis identified 44 compounds in the total extract and 33 compounds in the petroleum ether fraction of Botrychium ternatum. Conclusion: Botrychium ternatum demonstrated the best activity against M.sm among the 15 ethnomedicines. Further evaluation confirmed its significant anti-tuberculosis activity, particularly for its petroleum ether fraction. This study not only verified the feasibility of screening anti-tuberculosis agents from ethnomedicines, but also provided important candidate substances and scientific basis for developing new anti-tuberculosis drugs through compositional analysis.

Key words: Mycobacterium tuberculosis, Drug screening assays, Minimum inhibitory concentration, Ethnomedicines, Compositional analysis, Botrychium ternatum

中图分类号:

李静, 陈双双, 陈春华, 杨国祥, 杨承闹, 郑禄云, 谭承建, 李传友, 李洁, 周培富, 代小伟. 民族药阴地蕨抗结核活性筛选及化学成分分析[J]. 中国防痨杂志, 2026, 48(6): 787-794. doi: 10.19982/j.issn.1000-6621.20260021

Li Jing, Chen Shuangshuang, Chen Chunhua, Yang Guoxiang, Yang Chengnao, Zheng Luyun, Tan Chengjian, Li Chuanyou, Li Jie, Zhou Peifu, Dai Xiaowei. Screening of anti-tuberculosis activity and analysis of chemical constituents of the Ethnomedicine Botrychium ternatum[J]. Chinese Journal of Antituberculosis, 2026, 48(6): 787-794. doi: 10.19982/j.issn.1000-6621.20260021

图/表 5

表1

15种民族药治疗结核病应用记载

药物	民族药专著	应用记载	药用部位	使用民族
果上叶	《水族医药》^[24]	单方治疗肺结核	全草	彝、水、土家
	《楚雄彝州本草》^[25]	单方治疗肺结核
	《苗族医学》^[26]	肺结核(方剂:文殊兰、麦冬、岩豇豆、果上叶)
夏枯草	《苗族药物集》^[27]	复方治疗颈淋巴结结核(方剂:夏枯草、鱼腥草、一把伞、麦冬、八角枫、茜草)	全草	苗、壮
兔耳草	《苗族药物学》^[28]	单方治疗肺痨咯血	根	彝
何首乌	《土家族医药》^[29]	复方治疗淋巴结结核(方剂:马桑树皮,当归,何首乌)	块根	土家
金挖耳	《怒江流域民族医药》^[30]	单方治疗淋巴结结核	全草及根	苗
万寿竹	《苗族药物学》^[28]	单方治疗肺痨咳嗽	根或全草	苗
算盘子根	《苗族医学》^[26]	复方治疗颈淋巴结核(方剂:算盘子、岩豇豆、夏枯草、百部)	根或果实	苗
漆姑草	《苗族医学》^[26]	复方治疗肺结核(方剂:漆姑草、白及、岩豇豆、夏枯草)	全草	苗
侧柏叶	《德昂族药集》^[31]	复方治疗肺结核咯血(方剂:鲜仙鹤草、鲜旱莲草、侧柏叶)	干燥枝梢及叶	壮、德昂
	《简明壮医药学》^[32]	复方治疗肺结核(方剂:侧梢叶、生地、元参、百合、川贝、甘草)
桔梗	《中国苗族药物彩色图集》^[33]	复方治疗肺结核(方剂:桔梗、白及)	根	侗
商陆	《侗族药物方剂学》^[34]	单方治疗颈淋巴结结核	根	侗
小蓟	《苗族药物学》^[28]	肺痨咳嗽(煨水服或烧灰存性研粉吞服)	全草	侗
	《侗族药物方剂学》^[34]	单方对人型结核分枝杆菌有抑制作用
田基黄	《侗族医学》^[35]	治疗牛型结核分枝杆菌	全草	苗
射干	《侗族药物方剂学》^[34]	治疗结核病	根茎	侗、苗
阴地蕨	《苗族医学》^[26]	(1)复方治疗血虚痨病(方剂1:穿破石、鹿衔草、地骨皮、鳖甲、阴地蕨;方剂2:棉花根、黄芪、黄精、山楂、薏米、阴地蕨);	全草	彝、水、土家、苗、壮、侗、毛南
		(2)复方治疗小儿肺结核(方剂1:鱼腥草(白折耳)、凉粉柴、阴地蕨、猫爪草;方剂2:沙参、百合、小芝麻、麦冬、阴地蕨)
	《土家族医药》^[29]	复方治疗肺结核(方剂:百部、白及、白蔹、白贝七、白气参、三七、阴地蕨、百合、藕节、葎草花、肺心草、鱼鳅串、白奶参、阎王七)

表1

表2

表3

表4

图1

参考文献 48

[1]	屈燕, 李涛, 马文斌, 等. 世界卫生组织《2025年全球结核病报告》解读. 结核与肺部疾病杂志, 2025, 6(6): 613-623. doi:10.19983/j.issn.2096-8493.20250178.
[2]	Alghamdi S, Rehman SU, Shesha NT, et al. Promising Lead Compounds in the Development of Potential Clinical Drug Candidate for Drug-Resistant Tuberculosis. Molecules (Basel), 2020, 25(23):5685. doi:10.3390/molecules25235685.
[3]	Gopie FA, Commiesie E, Baldi S, et al. Should treatment of low-level rifampicin mono-resistant tuberculosis be different?. J Clin Tuberc Other Mycobact Dis, 2021, 23:100241. doi:10.1016/j.jctube.2021.100241.
[4]	Dhochak ND, Ramjat K M, Sankar JM, et al. Hypersensitivity Myocarditis Due to Antitubercular Drugs in Children. Pediatr Infect Dis J, 2020, 39(12): 1088-1091. doi:10.1097/inf.0000000000002844.
[5]	李维, 周志超, 郑洁, 等. 1995—2023年中国耐多药结核病研究的态势和热点分析. 中国防痨杂志, 2025, 47(6): 785-791. doi:10.19982/j.issn.1000-6621.20250112.
[6]	徐玉辉, 孙照刚. 抗结核药用植物的种类特点及其国内外比较. 中华中医药杂志, 2021, 36(3): 1582-1585.
[7]	Xu Y, Liang B, Kong C, et al. Traditional Medicinal Plants as a Source of Antituberculosis Drugs: A System Review. Biomed Res Int, 2021, 2021:9910365. doi:10.1155/2021/9910365.
[8]	Gautam S, Qureshi KA, Pasha SBJ, et al. Medicinal Plants as Therapeutic Alternatives to Combat Mycobacterium tuberculosis: A Comprehensive Review. Antibiotics (Basel), 2023, 12(3):541. doi:10.3390/antibiotics12030541.
[9]	Chen M, Liu J, Zou S, et al. A review on the ethnopharmacology, metabolites, pharmacological uses, and toxicology of Ficus hirta (Moraceae) Vahl. Front Pharmacol, 2025, 16:1545348. doi:10.3389/fphar.2025.1545348.
[10]	娄华勇, 傅建, 潘卫东. 基于UGM靶标的贵州药用植物抗结核分枝杆菌化合物构建及类似物研究. 贵州医科大学学报, 2023, 48(6):621-626,633. doi:10.19367/j.cnki.2096-8388.2023.06.001.
[11]	付宝慧, 张静. 冬虫夏草菌联合抗结核治疗肺结核的效果观察. 中国防痨杂志, 2024, (S2): 52-54.
[12]	Duan LY, Liang Y, Gong WP, et al. Comparative study on the antituberculous effect and mechanism of the traditional Chinese medicines NiuBeiXiaoHe extract and JieHeWan. Mil Med Res, 2021, 8(1): 34. doi:10.1186/s40779-021-00324-5.
[13]	Zhang X, Chen W, Du Y, et al. Phytochemistry and pharmacological activities of Arundina graminifolia (D.Don) Hochr. and other common Orchidaceae medicinal plants. J Ethnopharmacol, 2021, 276:114143. doi:10.1016/j.jep.2021.114143.
[14]	Fan Y, Wang M, Zhang Q, et al. Traditional uses, phytochemistry, pharmacology, toxicity and clinical application of traditional Chinese medicine Cynoglossum amabile: a review. Front Pharmacol, 2024, 15:1325283. doi:10.3389/fphar.2024.1325283.
[15]	Khan SA, Rather MA, Jia Z, et al. Discovery of antitubercular potential of trans-3-indoleacrylic acid and its derivatives targeting Mycobacterium tuberculosis: A combined in vitro and in silico investigation. Bioorg Chem, 2025, 163:108668. doi:10.1016/j.bioorg.2025.108668.
[16]	于兰, 陈双双, 王嫩寒, 等. 利福平耐药结核分枝杆菌对氟喹诺酮类药物表型耐药与其基因突变的一致性研究. 中国防痨杂志, 2024, 46(8): 942-950. doi:10.19982/j.issn.1000-6621.20240133.
[17]	刘瑞霞, 李翔辉, 冯青松. 乙醇回流法提取大别山产青钱柳总黄酮的工艺研究. 现代食品, 2022, 28(15): 77-79,83. doi:10.16736/j.cnki.cn41-1434/ts.2022.15.017.
[18]	王思雨, 张国玉, 张永祥, 等. 迷迭香粗提物提取工艺优化及其抑菌活性. 食品研究与开发, 2024, 45(9): 118-126. doi:10.12161/j.issn.1005-6521.2024.09.017.
[19]	CLSl. Performance standards for Antimicrobial susceptibility Testing. 36th ed. CLSl supplement M100. Clinical and Laboratory Standards Institute, 2026.
[20]	CLSl. Methods for Dilution Antimicrobial susceptibility Tests for Bacteria That Grow Aerobically 12th ed. CLSl standard M07 Clinical and Laboratory Standards Institute, 2024.
[21]	CLSI. Performance Standards for Susceptibility Testing of Mycobacteria, Nocardia spp., and Other Aerobic Actinomycetes. 2nd edition. CLSI supplement M24S. Clinical and Laboratory Standards Institute, 2023.
[22]	郭鸿, 么蕊, 范晶. 中草药寡糖的提取分离、结构鉴定及分析方法研究进展. 药物分析杂志, 2025, 45(3): 361-391. doi:10.16155/j.0254-1793.2024-1061.
[23]	何可群. HPLC-MS/MS法测定阴地蕨中山奈酚和槲皮素含量. 医药导报, 2015, (10): 1360-63. doi:10.3870/j.iss.1004-0781.2015.10.026.
[24]	王厚安. 水族医药. 贵阳: 贵州民族出版社, 1997.
[25]	王敏, 朱琚元. 楚雄彝州本草. 昆明: 云南人民出版社, 1998.
[26]	陆科闵, 王福荣. 苗族医学. 贵阳: 贵州科技出版社, 2006
[27]	陆科闵. 苗族药物集. 贵阳: 贵州人民出版社, 1988.
[28]	唐海华. 苗族药物学. 贵阳: 贵州民族出版社, 2006.
[29]	朱国豪. 土家族医药. 北京: 中医古籍出版社, 2006.
[30]	周元川, 郑进. 怒江流域民族医药. 昆明: 云南科技出版社, 2010.
[31]	方茂琴. 德昂族药集. 德宏: 德宏民族出版社, 2014.
[32]	钟鸣. 简明壮医药学. 南宁: 广西民族出版社, 2009.
[33]	汪毅. 中国苗族药物彩色图集. 贵阳: 贵州科技出版社, 2002.
[34]	龙运光, 袁涛忠. 侗族药物方剂学. 贵阳: 贵州科技出版社, 2009.
[35]	陆科闵. 侗族医学. 贵阳: 贵州科技出版社, 1992.
[36]	World Health Organization. Meeting report of the WHO expert consultaion on the definition of extensively drug-resistant tuberculosis, 27-29 October 2020. Geneva: World Health Organization, 2021.
[37]	Krishnaprasad VH, Nayak V, Kumar S. World Health Organisation’s Bacterial Pathogen Priority List (BPPL) 2017 and BPPL 2024 to combat global antimicrobial resistance crisis: ‘challenges and opportunities’. J Antimicrob Chemother, 2025, 80(8):2061-2069. doi:10.1093/jac/dkaf167.
[38]	Dvorakova M, Soudek P, Pavicic A, et al. The traditional utilization, biological activity and chemical composition of edible fern species. J Ethnopharmacol, 2024, 324:117818. doi:10.1016/j.jep.2024.117818.
[39]	Baran M, Grimes KD, Sibbald PA, et al. Development of small-molecule inhibitors of fatty acyl-AMP and fatty acyl-CoA ligases in Mycobacterium tuberculosis. Eur J Med Chem, 2020, 201: 112408. doi:10.1016/j.ejmech.2020.112408.
[40]	Niveditha N, Begum M, Prathibha D, et al. Design, Synthesis and Pharmacological Evaluation of Some C 3 Heterocyclic-Substituted Ciprofloxacin Derivatives as Chimeric Antitubercular Agents. Chem Pharm Bull (Tokyo), 2020, 68(12): 1170-1177. doi:10.1248/cpb.c20-00525.
[41]	Suryawanshi AG, Pathak C, Khona P, et al. Synthesis and biological evaluation of furan-1,3,4-oxadiazole as antitubercular, antibacterial, and antioxidant agents. Future Med Chem, 2025, 17(21): 1-16. doi:10.1080/17568919.2025.2570972.
[42]	Preez D Charné, Legoabe, et al. Arylnitro monocarbonyl curcumin analogues: Synthesis and in vitro antitubercular evaluation. Chem Biol Drug Des, 2023, 101(3): 717-726. doi:10.1111/cbdd.14174.
[43]	Montana M, Montero V, Khoumeri O, et al. Quinoxaline moiety: A potential scaffold against mycobacterium tuberculosis. Molecules, 2021, 26(16): 4742. doi:10.3390/molecules26164742.
[44]	Sonwane PN, Kumbhare MR. Molecular docking and pharmacokinetics of benzimidazole-based FtsZ inhibitors for tuberculosis. Sci Rep, 2025, 15(1): 35270. doi:10.1038/s41598-025-18084-w.
[45]	Chowdhury A, Bandyopanhyay A. Compelling cyclic peptide scaffolds for antitubercular action: An account (2011-21) of the natural source. Curr Protein Pept Sci, 2022, 23(12): 823-836. doi:10.2174/1389203723666220930111259.
[46]	Johansen MD Shalini, Sumit K, et al. Biological and biochemical evaluation of isatin-isoniazid hybrids as bactericidal candidates against Mycobacterium tuberculosis. Antimicrob Agents Chemother, 2021, 65(8): e0001121. doi:10.1128/aac.00011-21.
[47]	Barua Nilakshi, Buragohain, et al. Therapeutic Potential of Curcumin as an Antimycobacterial Agent. Biomolecules, 2021, 11(9):1278. doi:10.3390/biom11091278.
[48]	Sasikumar K, Ghosh AR, Dusthackeer A. Antimycobacterial potentials of quercetin and rutin against Mycobacterium tuberculosis H37Rv. Biotech, 2018, 8(10): 427. doi:10.1007/s13205-018-1450-5.

民族药阴地蕨抗结核活性筛选及化学成分分析

Screening of anti-tuberculosis activity and analysis of chemical constituents of the Ethnomedicine Botrychium ternatum

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药物	抑菌圈直径 (mm,$\overline{x}$±s)	药物	抑菌圈直径 (mm,$\overline{x}$±s)
果上叶	9±1.32	侧柏叶	9.25±1.00
夏枯草	9.3±0.76	田基黄	14.16±1.26
兔耳草	8.5±0.87	桔梗	-
何首乌	9.1±0.30	商陆	-
金挖耳	9.25±1.00	小蓟	-
万寿竹	-	射干	13.16±0.76
算盘子根	-	漆姑草	8.5±0.87
阴地蕨	18.33±4.04

提取物	M.sm	BCG
阴地蕨总浸膏	1875	2
石油醚相分离物	1875	1
二氯甲烷相分离物	1875	8
正丁醇相分离物	7500	16
利福平	0.5	0.015

药物	MDR-TB					RR-TB	Hr-TB	pre-XDR-TB	敏感菌株
药物	25-706	25-804	25-321	25-461	25-198	25-412	25-326	25-744	25-852	25-857	25-859
阴地蕨总浸膏	29	29	58	58	29	58	58	117	29	58	58
石油醚相分离物	14	14	29	29	14	29	29	58	14	29	29
利福平	≥64	≥64	≥64	≥64	1	≥64	0.25	≥64	0.06	0.03	0.03

[1]	王晓宁,何天伦,耿梦杰,宋渝丹,赵飞,何广学. 我国12省结核病门诊和实验室感染控制管理措施实施现状的抽样调查及其主成分分析[J]. 中国防痨杂志, 2018, 40(5): 519-524.
[2]	韩喜琴, 黄海荣, 李艳静, 于霞, 毕志强, 初乃惠, 马玙. 利福平和利福布汀对结核分枝杆菌交叉耐药率的初步探讨[J]. 中国防痨杂志, 2010, 32(11): 740-743.
[3]	高凌玉,张健源,李传友,周建琴,游雪甫,王振,汪月,孙承航,. 康乐霉素A体外抗结核活性的初步研究[J]. 中国防痨杂志, 2007, 29(2): 160-162.
[4]	吴风霞,李慧灵,王鑫君,. 结核分枝杆菌L型检测的临床意义[J]. 中国防痨杂志, 2004, 26(2): 107-109.