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香芹酚和普通百里香精油抑制生物膜形成,并与抗真菌药物协同作用,对耐药白念珠菌和热带念珠菌菌株有抑制作用。

Thymus vulgaris essential oil and thymol inhibit biofilms and interact synergistically with antifungal drugs against drug resistant strains of Candida albicans and Candida tropicalis.

机构信息

Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 2020022, India.

Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 2020022, India.

出版信息

J Mycol Med. 2020 Apr;30(1):100911. doi: 10.1016/j.mycmed.2019.100911. Epub 2019 Nov 7.

Abstract

Role of biofilm in disease development and enhance tolerance to antifungal drugs among Candida species has necessitated search for new anti-fungal treatment strategy. Interference in pathogenic biofilm development by new antifungal compounds is considered as an attractive anti-infective strategy. Therefore, the objective of this study was to evaluate Thymus vulgaris essential oil and its major active compound, thymol for their potential to inhibit and eradicate biofilms alone and in combination with antifungal drugs against Candida spp. with especial reference to Candida tropicalis. Anti-candidal efficacy of T. vulgaris and thymol in terms of minimum inhibitory concentration (MIC) was first determined to select the sub-MICs against C. albicans and C. tropicalis. Biofilm formation in the presence and absence of test agents was determined in 96-well microtiter plate by XTT reduction assay and effect of essential oils at sub-MICs of the test agents on biofilm development on glass surface was analysed by light and scanning electron microscopy. Synergistic interaction between essential oils and antifungal drugs were studied by checkerboard method. Effect of sub-MIC of T. vulgaris (0.5×MIC) and thymol (0.5×MIC) on biofilm formation showed a significant reduction (P<0.05) in biofilms. Light microscopy and SEM studies revealed disaggregation and deformed shape of C. albicans biofilm cells and reduced hyphae formation in C. tropicalis biofilm cells at sub-MICs of thymol. Significant effect of T. vulgaris and thymol was also recorded on pre-formed biofilms of both C. albicans and C. tropicalis. T. vulgaris and thymol also showed synergy with fluconazole against both in planktonic and biofilm mode of growth of C. albicans and C. tropicalis. However, synergy with amphotericin B is clearly evident only in planktonic Candida cells. Thyme oil and thymol alone or in combination with antifungal drugs can act as promising antibiofilm agent against drug resistant strains of Candida species and needs further in vivo study to synergise its therapeutic efficacy.

摘要

生物膜在疾病发展中的作用以及增强了念珠菌属对抗真菌药物的耐受性,这使得人们必须寻找新的抗真菌治疗策略。通过新的抗真菌化合物干扰致病性生物膜的形成被认为是一种有吸引力的抗感染策略。因此,本研究的目的是评估香芹酚及其主要活性化合物麝香草酚单独使用和与抗真菌药物联合使用对念珠菌属(尤其是热带念珠菌)生物膜的抑制和清除作用。首先,根据最低抑菌浓度(MIC)确定了香芹酚和麝香草酚的抗真菌功效,以选择针对白色念珠菌和热带念珠菌的亚 MIC 值。通过 XTT 还原测定法在 96 孔微量滴定板中确定了在存在和不存在测试剂的情况下生物膜的形成,并且通过光和扫描电子显微镜分析了测试剂的亚 MIC 对玻璃表面生物膜发育的影响。通过棋盘法研究了精油与抗真菌药物之间的协同作用。香芹酚(0.5×MIC)和麝香草酚(0.5×MIC)的亚 MIC 对生物膜形成的影响显示生物膜的形成显著减少(P<0.05)。光显微镜和 SEM 研究显示,在麝香草酚的亚 MIC 下,白色念珠菌生物膜细胞解聚和变形,热带念珠菌生物膜细胞中菌丝形成减少。香芹酚和麝香草酚对两种念珠菌的预形成生物膜也有显著影响。香芹酚和麝香草酚还显示与氟康唑联合使用对白色念珠菌和热带念珠菌的浮游和生物膜生长模式具有协同作用。然而,与两性霉素 B 的协同作用仅在浮游念珠菌细胞中明显。单独使用或与抗真菌药物联合使用的香芹酚和麝香草酚可以作为有前途的抗生物膜剂,用于对抗耐药的念珠菌属菌株,并且需要进一步的体内研究来协同其治疗效果。

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