摘要
目的探讨大蒜素在体外对于白色念珠菌生物膜作用的抗击作用。方法通过用4-2 048μg/ml二倍倍比稀释的大蒜素或0. 125-1 024μg/ml二倍倍比稀释的氟康唑处理白色念珠菌48 h,24 h或8 h后,采用XTT法检测大蒜素对白色念珠菌的杀伤作用,以及大蒜素造成生物膜80%抑制浓度(SMIC_(80))的测定,采用FUNI免疫荧光染色的方式测定大蒜素对白色念珠菌形成的生物膜厚度的影响,采用q PCR检测大蒜素对白色念珠菌中与生物膜形成相关的基因表达水平。结果大蒜素对于所有白色念珠菌的MIC在32-128μg/ml,SMIC_(80)为64-256μg/ml,均小于对应到氟康唑的作用值。大蒜素对于白色念珠菌标准株SC5314的生物膜形成呈现剂量依赖的抑制效果,当大蒜素浓度为4μg/ml时对生物膜形成的抑制作用不明显,随着剂量的升高开始表现出抑制作用,而当大蒜素浓度升为128μg/ml时,对生物膜形成的抑制作用达到80%,显著高于对照组(大蒜素浓度为0)(P <0. 001);当大蒜素的浓度为16μg/ml及以下时,对于菌株的抑制作用不明显(P> 0. 05),而当大蒜素的浓度升高到128μg/ml时,对菌株的抑制作用表现明显(P <0. 01),并且在第4小时时lg CFU降为4。通过对染色后的荧光强度进行量化来显示生物膜的厚度,可以发现,荧光强度随着大蒜素的浓度升高而不断降低,在大蒜素浓度为128μg/ml时,荧光强度值接近于0。对生物膜形成相关基因表达情况进行q PCR测定,发现经过大蒜素128μg/ml处理8 h后,相比不处理,生物膜相关基因ALS1表达下调了4. 7%,SUN41下调了18. 7%,MP65下调了68. 3%,HWP1下调了77. 4%,GSH下调了38. 4%。结论大蒜素对于白色念珠菌具有抑制作用,并对其表面生物膜的形成具有良好的抵抗作用,效果显著,值得临床参考。
Objective To investigate the effect of allicin against the biofilm of Candida albicans in vitro . Methods Candida albicans was treated with 4-2 048 μg/ml double diluted allicin or 0.125-1 024 μg/ml double diluted fluconazole for 48 h, 24 h or 8 h, respectively, and then the killing effect of allicin and 80% inhibitory concentration on biofilm(SMIC 80 ) were detected by XTT assay, the effect of allicin on biofilm thickness formed by Candida albican was detected using FUNI immunofluorescent staining, and the expression levels of genes related with biofilm formation in Candida albican were detected by qPCR. Results Except the standard strain SC5314 and the strains other than the clinical strains No. 4 and No. 6, the MIC of fluconazole was above 128 μg/ml, which was defined as a drug-resistant strain, and the SMIC 80 of all strains for fluconazole was 1 024 μg/ml or more. The allicin had an MIC of 32- 128 μg/ml for all Candida albicans and 64-256 μg/ml for SMIC 80 , which were smaller than the corresponding values for fluconazole. Allicin had a dose-dependent inhibitory effect on the biofilm formation of Candida albicans SC5314. While the allicin concentration was 4 μg/ml, the inhibitory effect on biofilm formation was not obvious, and the inhibition was gradually increased with the increasing dose. When the concentration of allicin increased to 128 μg/ml, the inhibitation on biofilm formation reached 80%, which was significantly higher than that of controls(0 μg/ml, P <0.001). When the concentration of allicin was 16 μg/ml and below, the inhibitory effect on the strain was not significant( P >0.05). When the concentration of allicin was increased to 128 μg/ml, the inhibitory effect on the strain was significant( P <0.01), and the lgCFU decreased to 4 at 4 h. The thickness of the biofilm was quantified by the fluorescence intensity after staining and the fluorescence intensity decreased with the increasing concentration of allicin. When the allicin concentration was 128 μg/ml, the fluorescence intensity v
作者
符林瑜
林景
徐博譞
FU Linyu;LIN Jing;XU Boxuan(Department of Pharmacy,Fifth People’s Hospital of Hainan Province,Haikou 570206,China)
出处
《山西医科大学学报》
CAS
2019年第4期456-460,共5页
Journal of Shanxi Medical University
基金
海南省自然科学基金资助项目(817374)
