中药白头翁对体外大肠杆菌生物膜抑制作用探究.doc

中药白头翁对体外大肠杆菌生物膜抑制作用探究 　　[摘要] 目的 在Φ30培养皿和96孔板表面建立大肠杆菌生物被膜（E.coli biofilm，EBF），形成体外模型，并开展白头翁对EBF抑制作用的初步研究。 方法 选取临床分离的大肠杆菌菌株，在Φ30培养皿和96孔板中分别复制体外EBF模型，将白头翁作用于EBF体外模型，分别采用银染法和MTT法考察白头翁对EBF的影响。 结果 Φ30培养皿中，表面观察到黑色呈棉絮状的膜样物，空白组没有此样物质；96孔板中，模型组的OD值为0.079，空白组的OD值为1.548；白头翁醇提组抑制率为25.79%，白头翁水提组抑制率为53.11%；750μg/mL的白头翁对大肠杆菌生物被膜有最佳抑制作用，抑制率为55.17%；药物作用12h后开始白头翁组的EBF明显少于空白对照组，24h效果最好。 结论 Φ30培养皿和96孔板表面可以形成大肠杆菌生物被膜体外模型，白头翁可以抑制和破坏早期及成熟的细菌生物膜，且其抑制作用表现出了一定的量效关系。 [关键词] 大肠杆菌；生物被膜；MTT法；银染法；白头翁 [中图分类号] S853.74 [文献标识码] B [文章编号] 2095-0616（2013）24-20-03 Establishment of in vitro model for escherichia coli biofilm and preliminary study inhibitory effect of Pulsafilla on the biofilm YAO Yu1 BAO Yongrui1，2 MENG Xiansheng1，2 WANG Shuai1，2 1.Liaoning University of Traditional Chinese Medicine，Dalian 116600，China；2.Liaoning Modern Traditional Chinese Medicine Research and Engineering Laboratory，Dalian 116600，China [Abstract] Objective To establish in vitro model of escherichia coli biofilm（EBF） formated on Φ30 petri dishes and 96-well plates，and investigate the inhibitory effect of Pulsafilla on the EBF. Methods At the Φ30 petri dishes and 96-well plates formd the in vitro model of EBF，Use the inhibitory effect of Pulsafilla on EBF as the examination indicator，MTT assay was used to detect the best extraction solvent and the best inhibit rate，and drawed the growh curve；Silver staining method was used to observe the morphology of EBF in Φ30 petri dishes. Results There were black stained cotton wool membrane-like material appeared on the surface of Φ30 petri dishes with Escherichia coli，but the blank group didn’t have.In 96-well plates，the OD values of model group was 1.548，blank group was0.079；The inhibit effect of Pulsafilla water -extraction was 25.79%，The inhibit effect of Pulsafilla alcohol -extraction was 53.11%.When the concentration of Pulsafilla was 750μg/mL，the best inhibition rate was 55.17%.Pulsafilla acted on EBF 12 hours later，the EBF in drug group was less than that of bl