中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (4): 262-265.doi: 10.3760/cma.j.issn.0412-4030.2019.04.008

• 研究报道 • 上一篇    下一篇

蛇床子素体外对酵母相马尔尼菲篮状菌的抗菌活性检测

罗秋红1    潘开素2    罗宏2    曹存巍2   

  1. 1广西中医药大学第一附属医院皮肤科，南宁  530023； 2广西医科大学第一附属医院皮肤科，南宁  530021
  • 收稿日期:2018-07-31 修回日期:2018-12-28 发布日期:2019-04-01
  • 通讯作者: 曹存巍 E-mail:caocunwei@yeah.net
  • 基金资助:
    国家自然科学基金（81571971、81271804）；广西壮族自治区卫生厅计划课题（Z2013165）

Evaluation of in vitro antifungal activity of osthole against Talaromyces marneffei in yeast phase

Luo Qiuhong1, Pan Kaisu2, Luo Hong2, Cao Cunwei2   

  1. 1Department of Dermatology, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, China;  2Department of Dermatology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China
  • Received:2018-07-31 Revised:2018-12-28 Published:2019-04-01
  • Contact: Cao Cunwei E-mail:caocunwei@yeah.net
  • Supported by:
    National Natural Science Foundation of China （81571971, 81271804）; Project of Health Department of Guangxi Zhuang Autonomous Region （Z2013165）

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摘要： 目的 体外检测蛇床子素对酵母相马尔尼菲篮状菌（Talaromyces marneffei，TM）的抗菌活性，为临床用中药治疗马尔尼菲篮状菌感染提供实验依据。方法 TM菌株20株，包括1株标准株， 2株氟康唑天然耐药株，11株临床分离株，6株野生竹鼠分离株。使用96孔细胞培养板，采用微量稀释法制备蛇床子素、氟康唑、两性霉素B、伊曲康唑、伏立康唑药敏板。将TM受试菌以（1～5） × 103 CFU/ml菌悬液接种于药敏板，在 37 ℃下培养48 h后观察结果，同时设阳性对照（仅加入TM和培养基，不加药物）和阴性对照（仅加入培养基，不加TM和药物）。参照美国临床实验室标准研究所颁布的酵母菌抗真菌药物敏感性试验方案 M27-A2判定最低抑菌浓度（MIC值）。与生长对照孔相比， ≥ 80%生长抑制所对应的最低药物浓度为氟康唑MIC，其余药物MIC为100%生长抑制所对应的最低药物浓度。结果 质控株MIC值均在参考范围内，阳性对照孔TM生长良好。氟康唑MIC值范围2.0～8.0 mg/L（对氟康唑天然耐药株为128 mg/L），两性霉素B 1.0～4.0 mg/L，伊曲康唑0.03～0.25 mg/L，伏立康唑0.06～0.25 mg/L。蛇床子素对标准株FRR2161的MIC为16 mg/L，对氟康唑天然耐药株的MIC均为32 mg/L，对1株野生竹鼠分离株MIC为128 mg/L，对其他16株TM菌株的MIC值范围为16～64 mg/L，MIC90及MIC50分别为64和32 mg/L。结论 蛇床子素具有抑制酵母相马尔尼菲篮状菌的活性。

关键词: 踝节菌属, 蛇床子素, 微生物敏感性试验, 马尔尼菲篮状菌

Abstract: 【Abstract】 Objective To evaluate the in vitro antifungal activity of osthole against Talaromyces marneffei （TM） in yeast phase, in order to provide an experimental reference for the clinical treatment of TM infection with Chinese medicine. Methods There were 20 TM strains, including 1 standard strain, 2 fluconazole-spontaneously resistant strains, 11 clinical isolates, and 6 isolates from wild bamboo rats. A microdilution method was used to prepare 96-well antifungal sensitivity test plates containing osthole, fluconazole, amphotericin B, itraconazole and voriconazole at different concentrations, which were incubated with （1 - 5） × 103 CFU/ml of tested TM strain suspensions at 37 ℃ for 48 hours. Meanwhile, TM strains cultured in the media without antifungal drugs served as positive （growth） control group, and culture media served as negative group. The minimum inhibitory concentrations （MICs） of antifungal drugs against yeasts were determined using the Clinical and Laboratory Standards Institute （CLSI） broth microdilution susceptibility method （M27-A2 Document）. Fluconazole MIC was defined as the lowest drug concentration that resulted in ≥ 80% growth inhibition, and MICs of other antifungal drugs were the lowest drug concentrations that resulted in 100% growth inhibition, compared with growth control wells. Results The MICs among the quality control strains were all within the reference range, and TM grew well in the positive control wells. The MIC ranges of fluconazole, amphotericin B, itraconazole and voriconazole against TM strains were 2.0 - 8.0 mg/L, 1.0 - 4.0 mg/L, 0.03 - 0.25 mg/L and 0.06 - 0.25 mg/L respectively, and the MIC of fluconazole against fluconazole-spontaneously resistant strains was 128 mg/L. The MICs of osthole against the TM standard strain （FRR2161）, fluconazole-spontaneously resistant strains and 1 isolate from wild bamboo rats were 16, 32 and 128 mg/L respectively, and the MIC range of osthole against other 16 TM strains was 16 - 64 mg/L. The MICs of osthole at which 90% and 50% of the TM strains were inhibited were 64 and 32 mg/L respectively. Conclusion Osthole exhibits the antifungal activity against the yeast form of TM.

Key words: Talaromyces, Osthole, Microbial sensitivity tests, Talaromyces marneffei

引用本文

罗秋红 潘开素 罗宏 曹存巍. 蛇床子素体外对酵母相马尔尼菲篮状菌的抗菌活性检测[J]. 中华皮肤科杂志, 2019,52(4):262-265. doi:10.3760/cma.j.issn.0412-4030.2019.04.008

Luo Qiuhong, Pan Kaisu, Luo Hong, Cao Cunwei. Evaluation of in vitro antifungal activity of osthole against Talaromyces marneffei in yeast phase[J]. Chinese Journal of Dermatology, 2019, 52(4): 262-265.doi:10.3760/cma.j.issn.0412-4030.2019.04.008