荷叶提取物抑制粪肠球菌浮游细菌及其生物膜  

Lotus Leaves Extracts Inhibits Planktonic Enterococcus faecalis and Its Biofilm

作　　者：潘利君 胡翔宇 李宗莉 赵瑜[1] 邓黎超 潘乙怀[1] PAN Li-jun;HU Xiang-yu;LI Zong-li;ZHAO Yu;DENG Li-chao;PAN Yi-huai(Department of Dentistry and Endodontics,Stomatological Hospital Affiliated of Wenzhou Medical University,Zhejiang 325027)

机构地区：[1]温州医科大学附属口腔医院牙体牙髓科,浙江325027

出　　处：《中国中西医结合杂志》2025年第3期337-342,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：温州市重大科技创新攻关项目(No.ZY2022019);浙江省基础公益研究计划项目(No.LGF21H140005)。

摘　　要：目的探究荷叶提取物对粪肠球菌浮游细菌及其生物膜的影响。方法制备荷叶水提取物、荷叶乙醇提取物、荷叶总黄酮提取物及荷叶生物碱粗提取物。采用二倍梯度稀释法及平板涂布法测定各荷叶提取物对粪肠球菌的最小抑菌浓度(MIC);随后根据结果分为1.3、0.65 g/mL荷叶生物碱粗提取物组,阳性对照组(2%氯己定溶液)及阴性对照组(无菌PBS溶液)。采用死菌/活菌荧光染色剂,并通过激光扫描共聚焦显微镜观察经荷叶生物碱粗提取物处理后粪肠球菌生物膜的形成情况;采用吖啶橙/溴化乙锭染色,荧光显微镜下观察经荷叶生物碱粗提取物处理后粪肠球菌生物膜结构中的死活菌情况。结果荷叶水、乙醇提取物及荷叶总黄酮、生物碱粗提取物对粪肠球菌的MIC分别为7.04、1.76、>1.3、1.3 g/mL。在粪肠球菌生物膜形成及已形成的生物膜结构中,与阴性对照组比较,1.3、0.65 g/mL荷叶生物碱粗提取物组及阳性对照组,活菌百分比均减少(P<0.01)。与阳性对照组比较,0.65 g/mL荷叶生物碱粗提取物活菌百分比增加(P<0.01),而1.3 g/mL荷叶生物碱粗提取物则差异无统计学意义(P>0.05)。结论荷叶生物碱粗提取物能有效抑制粪肠球菌生长及其生物膜的形成及结构,在根管治疗中具有良好的应用前景。Objective To investigate the effects of lotus leaf extracts on the planktonic bacteria and biofilm of Enterococcus faecalis.Methods Lotus leaf water extract,ethanol extract,total flavonoid extract,and crude alkaloid extract were prepared.The minimum inhibitory concentration(MIC)of each lotus leaf extract against Enterococcus faecalis was determined by two-fold gradient dilution and plate coating methods.Based on the results,the experimental groups were divided into 1.3 g/mL and 0.65 g/mL crude alkaloid extract groups,positive control group(2%chlorhexidine solution),and negative control group(sterile PBS solution).The LIVE/DEAD BacLight~(TM)Bacterial Viability Kit was used for fluorescence staining of live/dead bacteria,and the formation of Enterococcus faecalis biofilm after treatment with the crude alkaloid extract was observed using confocal laser scanning microscopy(CLSM).The structure of live and dead bacteria in the biofilm was observed under a fluorescence microscope after acridine orange/ethidium bromide staining.Results The MICs of lotus leaf water extract,ethanol extract,total flavonoid extract,and crude alkaloid extract against Enterococcus faecalis were 7.04,1.76,>1.3,and 1.3 g/mL,respectively.In the formation and structure of Enterococcus faecalis biofilm,compared with the negative control group,the percentage of live bacteria in the 1.3 g/mL and 0.65 g/mL crude alkaloid extract groups and the positive control group significantly reduced(P<0.01).Compared with the positive control group,the percentage of live bacteria in the 0.65 g/mL crude alkaloid extract group increased(P<0.01),while there was no statistically significant difference in the 1.3 g/mL crude alkaloid extract group(P>0.05).Conclusion The crude alkaloid extract of lotus leaves can effectively inhibit the growth of Enterococcus faecalis and the formation and structure of its biofilm,suggesting good potential for application in root canal therapy.

关 键 词：荷叶提取物 粪肠球菌 生物膜 死/活菌染色 中药提取物 

分 类 号：R285.5[医药卫生—中药学]