The Inhibitory Effect Of Matrine On Escherichia Coli Biofilm Formation In Vivo

Part I The inhibitory effect of different antimicrobial concentrations matrine on Escherichia coli biofilm for mation in vivoObject This experiment adopts the LB broth—peritoneal dialysis catheter system establishment of Escherichia coli biofilm model in vitro,To investigate the influence on the formation of Escherichia coli biofilm in vitro of matrine.Methods Frist,Using matrine and levofloxacin to measure the minimum inhibitory concentration(MIC)for Escherichia coli.The high pressure sterilized peritoneal dialysis tube carriers are inserted into the 24 well plate,Divided into 6 groups,blank control group,1/2 MIC of levofloxacin group(opstive control group),1/2MIC、1/4MIC、1/8MIC、1/16MIC matrine group.The establishment of Escherichia col i biofilm model from the beginning,will be drug intervention group addi ngdrugs to tune into the corresponding drug concentration.In 1 days,3 da y-s,7 days SEM and silver staining observe the surface of the dialysis ca th-eter tube’s BF,and crystal violet staining BF semi-quantitative,using fl at-panel microquan-titative method to count viable cells on the surface of the carrier.Results Matrine MIC 4096μg/ml,and levofloxacin MIC 0.015625 μg/ml.In both 1 and 3、7 days,Found that 1/2MIC matrine group and blank-gro-up comparedto the formation of caninhibit Escherichiacoli biofilm,compared withl/2MIC levofloxacin group had no statistical differ-ent-ce(P>0.05),The fastsilverstaining observation,1/2 MIC ofmatrine and 1/2MIC levofloxacin group carrier surface are decentralized,mass like black cotton wool structure is less than the blank group,other inhi-bitory concentration of matrine and the blank group were similar,but by scanning electronmicroscope and other inhibitory concentration of matrine though not on biofilm formation have much impact,but the impact of biofilm morphology after drug action,under scanning electron microsc-opy of biological membranes tructure is loose,The cells around mucin-ous substance less.Conclusion 1/2 MIC matrine and 1/2 MIC levofloxacin group in vitro biofilm formation of Escherichia coli inhibited,while other concentrateion of Matrine on Escherichia coli biofilmhad has no inhibitory effect,But under the scanning electron microscope(SEM)observation findings have effection for 1/4 MIC、1/8 MIC、1/16 MIC matrine group biofilm morphology.Part II The effect of different antimicrobial concentr ations of Matrine on the expression of outer membran e protein A mRNA in Escherichia coliObjective To investigate the different inhibitory concentration matrine effects in biological membrane to t he early,middle and late on the Escherichia coli outer membrane protein A(OmpA)mRNA expression.Method Established from the start of Escherichia coli biofilm model,drug intervention group add drug tune into the corresponding drug concentration.In Chapter 1,3,7 days from 24 orifice plate of total bacterial RNA.According to Takara kit was revers transcribed into cDNA,real-time fluorescence quantitative PCR observation at the same time,different concentration of Escherichia coli outer membrane protein A(OmpA)mRNA expression.Result For the Escherichia coli OmpA mRNA expression,in Escherichia coli biofilm formation of early,middle and late 1/2MIC levofloxacin group,1/2 MIC matrine group,1/4 MIC matrine group,1/8 MIC matrine group,1/16MIC Sophora flavescens alkali group compared with those in the control group compared to inhibit the expression of OmpA mRNA(P<0.05)and different period concentration of Matrine on the OmpA mRNA expression affects the amount of no statistical difference.(P>0.05)Conclusion 1/2 MIC matrine group,1/4MIC matrine group,1/8MIC matrine group,1/16MIC matrine group and 1/2 MIC Levofloxacin group in the biofilm formation of early,middle and late can inhibit the OmpA mRNA expression.