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

doi:10.19982/j.issn.1000-6621.20260021

Abstract

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

CLC Number:

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

Figures/Tables 5

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)

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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