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- 2015
真菌密度感应分子Farnesol在表皮葡萄球菌和白假丝酵母菌混合生物膜形成中的作用
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Abstract:
摘要：目的 探讨真菌密度感应分子Farnesol在表皮葡萄球菌和白假丝酵母菌混合生物膜形成中的作用。方法 将表皮葡萄球菌标准株ATCC35984及白假丝酵母菌标准株ATCC10231混合培养建立体外生物膜模型，加入Farnesol或不加入Farnesol分为Farnesol处理组和对照组，孵育2、4、6、8、12、24、48、72、96h时用结晶紫染色法半定量检测生物膜形成能力；二甲氧唑黄(XTT)比色法评价生物膜的体外生长动力学；扫描电子显微镜观察生物膜超微结构；荧光定量PCR分析各组生物膜形成相关基因的表达情况。结果 结晶紫染色法生物膜半定量检测显示，12、24h Farnesol处理组和对照组差异有统计学意义(P<0.05)。XTT比色法生长动力学检测显示，Farnesol处理组在4、48h与对照组比较，差异有统计学意义(P<0.05)。扫描电镜结果显示，对照组混合生物膜结构较Farnesol处理组致密复杂。荧光定量PCR结果显示，培养6h Farnesol处理组表皮葡萄球菌生物膜形成相关基因icaA、fbe、aap基因及白假丝酵母菌生物膜形成相关基因als3、hwp1、efg1基因的表达下调；培养24h Farnesol处理组表皮葡萄球菌生物膜形成相关基因aap基因及白假丝酵母菌生物膜形成相关基因als3、hwp1、efg1基因的表达下调。结论 在真菌密度感应分子Farnesol的干预下，对照组形成的混合生物膜较Farnesol处理组结构更为致密及复杂，这种结构的改变可能与Farnesol处理组白假丝酵母菌生物膜形成相关基因als3、hwp1、efg1基因的表达下调相关性更密切。
ABSTRACT: Objective To explore the function of Farnesol in the formation of Staphylococcus epidermidis-Candida albicans mixed species biofilms. Methods The strains of Staphylococcus epidermidis ATCC35984 and Candida albicans ATCC10231 were co-cultured to form mixed species of biofilms. Then they were divided into Farnesol group and control group based on treatment with Farnesol or not. Biofilm mass was semi-quantified by crystal violet semi-quantitative adherence assay at 2, 4, 6, 8, 12, 24, 48, 72 and 96h of incubation. XTT assay was performed to determine the growth kinetics at the same time. Scanning electron microscopy was used to observe the ultrastructure of the biofilms. Real-time PCR was used to analyze the expressions of icaA, fbe, aap, hwp1, als3 and efg1 genes. Results Crystal violet semi-quantitative adherence assay showed that at 12 and 24h of incubation, Farnesol group had significantly decreased optical density compared to control group (P<0.05). XTT assay showed that at 4 and 48h of incubation, biofilms growth kinetics differed significantly in Farnesol group and control group(P<0.05). Scanning electron microscopy showed that the three-dimensional structure of mix-biofilm in control group was more complex. As real-time PCR showed, after 6-hour culture, the expressions of icaA, fbe, aap, hwp1, als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group and after 24-hour culture the expressions of aap, hwp1, als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group. Conclusion With the intervention by fungal quorum sensing molecules Farnesol, the structure in control group became denser and more complex than that in Farnesol group. This change may have a closer correlation with the down-regulated expressions of als3, hwp1 and efg1 genes, which are related to
| [1] | 龚凤云，刘丽娜，邢铭友，等.表皮葡萄球菌耐药与生物膜的相关研究[J]. 中华医院感染学杂志, 2011, 21(1):20-23. |
| [2] | ZIEBUHR W, HENNIG S, ECKART M et al. Nosocomial infections by Staphylococcus epidermidis: how a commensal bacterium turns into a pathogen[J]. Int J Antimicrob Agents, 2006, 28(Suppl 1):S14-S20. |
| [3] | 李燕，李冬冬，陶传敏，等. 表皮葡萄球菌生物膜形成及相关基因的检测与评价[J]. 中华医院感染学杂志, 2010, 20(4):473-476. |
| [4] | CHEN L, WEN YM. The role of bacterial biofilm in persistent infection and control strategies[J]. Int J Oral Sci, 2011, 3(2):66-73. |
| [5] | PARK SJ, HAN KH, PARK JY, et al. Influence of bacterial presence on biofilm formation of Candida albicans[J]. Yonsei Med J, 2014, 55(2):449-458. |
| [6] | CERCA N, GOMES F, PEREIRA S, et al. Confocal laser scanning microscopy analysis of S. epidermidis biofilms exposed to farnesol, vancomycin and rifampicin[J]. BMC Research Notes, 2012, 5:244-250. |
| [7] | 丁进亚，黄前川，徐娟，等. 置入性导管相关感染的病原菌与生物被膜形成[J]. 医学综述, 2012,18(6):933-935. |
| [8] | SUTTER D, STAGLIANO D, BRAUN L, et al. Polymicrobial bloodstream infection in pediatric patients: risk factors, microbiology, and antimicrobial management[J]. Pediatr Infect Dis J, 2008, 13(5):400-405. |
| [9] | 汤琦，袁兵，黄云超，等. 乳腺外科表皮葡萄球菌icaA、icaD及聚集相关蛋白对生物膜形成的影响[J]. 中国修复重建外科杂志, 2014, 28(2):244-249. |
| [10] | ZHAO X, DANIELS KJ, OH SH, et al. Candida albicanas Als3 is required for wild-type biofilm formation on silicone slastomer surfaces[J]. Microbiology, 2006, 152(8):2287-2299. |
| [11] | NOBILE CJ, NETT JE, ANDES DR, et al. Function of Candida albicans adhesin Hwp1 in biofilm formation[J]. Eykaryot Cell, 2006, 5(10):1604-1610. |
| [12] | 娄强，王艳歌，瞿涤. 表皮葡萄球菌双组分信号转导系统saeRS对相关蛋白调控的研究[J]. 西安交通大学学报：医学版. 2012, 33(6):680-684. |
| [13] | 叶联华，黄云超，杨达宽，等. 表皮葡萄球菌ica操纵子与聚氯乙烯材料表面细菌生物膜形成的关系[J]. 中华医院感染学杂志, 2010, 20(24):3841-3843. |
| [14] | ALBUQUERQUE P, CASADEVALL A. Quorum sensing in fungi－a review[J]. Med Mycol, 2012, 50(4):337-345. |
| [15] | SHIRTLIFF ME, PETERS BM, JABRA-RIZK MA. Cross-kingdom interactions: Candida albicans and bacteria[J]. FEMS Microbiol Lett, 2009, 299(1):1-8. |
| [16] | FRANCA A, FREITAS AI, HENRIQUES AF, et al. Optimizing a qPCR gene expression quantification assay for S. epidermidis Biofilms: A comparison between commercial kits and a customized protocol[J]. PloS DNE, 2012, 7(5):e37480. |
| [17] | AGARWAL A, JAIN A. Glucose & chloride induce biofilm production & ica operon in clinic isolates of Staphylococci[J]. Indian J Med Res, 2013, 138(2):262-266. |
| [18] | 叶联华，黄云超，许赓，等. 医源性表皮葡萄球菌胞间粘附素基因操纵子在聚氯乙烯材料表面细菌生物膜形成中的作用研究[J]. 中国修复重建外科杂志, 2011, 25(4):466-471. |
| [19] | STEVENS NT, THARMABALA M, DILLANE T, et al. Biofilm and the role of the ica operon and aap in Staphylococcus epidermidis isolates causing neurosurgical meningitis[J] . Clin Microbiol Infect, 2008, 14(7):719-722. |
