齐墩果酸联合二甲亚砜对生物膜中粪肠球菌抑菌作用研究.docVIP

齐墩果酸联合二甲亚砚对生物膜中粪肠球菌抑菌作用研究 【摘要】目的：研究齐墩果酸联合二甲亚飒对 单一生物膜中粪肠球菌的抑菌效果。方法：（1）采用 微量棋盘液体稀释法测定齐墩果酸与二甲亚砚联合抑 菌浓度分数指数（FICl）o （2）建立24 h粪肠球菌生物 膜模型，随机分为三组：1 mg/mL齐墩果酸+50%二甲 亚砚（A组）、2%氯己定（B组）、去离子水（C组）。 各组药物分别作用30 min后荧光染色，激光共聚焦显 微镜（CLSM）观察染色情况及死、活菌数量变化。 结果：（1）二甲亚砚和齐墩果酸对粪肠球菌的 MIC分别 20%和 100 Pg/mLo联合时 FICI=0.5125o（2） A组、B组生物膜中以死菌为主，仅见散在零星分布 活菌，C组生物膜中以活菌为主，仅存在极少量死菌。 结论：齐墩果酸与二甲亚枫联合，对体外粪肠球 菌的抗菌作用表现为相加作用，1 mg/mL齐墩果酸联 合50%二甲亚飒可有效抑制生物膜中粪肠球菌生长。 【关键词】齐墩果酸；二甲亚砚；粪肠球菌; 激光共聚焦显微镜 [Abstract] Objective： To evaluate the antibacterial effect of oleanolic acid (OA) and dimethyl sulfoxide (DMSO) against enterococcus faecalis (E.faecalis) in biofilm in vitro.Method： (1) The microscale checkerboard technique was applied to calculate the FICI value of OA combinating with DMSO. (2) E.faecalis biofilm models were formed after 24h? Samples were randomly divided into three groups： 1 mg/mL OA+50% DMSO (group A), 2% chlorhexidine (group B), deion ized water (group C)? Medicati ng in each groups for 30 min respectively.After fluorescent staining, samples were observed by CLSM.Result： (1) The MICs of DMSO and OA against E.faecalis were 20% and 100 u g/mL.FICI=0.5125 when they were combined?(2) The biofilms of group A and B were mainly composed of dead bacteria, only scattered living bacteria could be found.The biofilm of group C was mainly composed of living bacteria, only tiny dead bacteria could be detected. Conclusion： Additive joint action was existed between OA and DMSO against E.faecalis.1 mg/mL OA combinating with 50% DMSO can effectively remove the bacteria in biofilms. 【Key words】 Oleanolic acid； Dimethyl sulfoxide； En terococcus faecalis； Con focal laser sea nning microscope First-author s address： Southwest Medical University, Luzhou 646000, China doi： 10.3969/j.issn.1674-4985.2016.14.003 粪肠球菌(Enterococcus faecalis, E.faecalis)具有 超强的生存能力，在根管内形成生物膜后，粪肠球菌 可明显增强抗力近1000倍，引起根管治疗后再感染, 或导致难治性根尖周炎的迁移不愈［1-4］o粪肠球菌耐 药性强，目前常用的根管消毒药物对于抑制根管内粪 肠球菌生长的有效性、安全性尚不能达到较为理想的 效果⑸。齐墩果酸(OleanolicAcid, 0A)是五环三菇 类化合物，普遍存在于多种天然植物中，毒