The Effect Of Matrine Against Biofilm Formation In Escherichia Coli In Vivo

Part I The effect of different concentrations of matrine on the biofilm formation in Escherichia coli: the regualtion of Omp AObject To investigate the effect of different concentrations of matrine on the expression of outer membrane protein A(Omp A)in the middle and later phases of biofilm formation of Escherichia coli.Methods Escherichia coli(ATCC25922)was used in this study to measure the minimum inhibitory concentration(MIC)of matrine and levofloxacin for it.And then biofilms was induced using several peritoneal dialysis catheters in 24-well plates and divided into 5 groups: blank control group(negative control),1MIC,2MIC,4MIC matrine groups and 1MIC of levofloxacin group(positive control group),after 3 days and 7 days of culturing(corresponding to the middle and later phases of biofilm formation).After treatment of Levofloxacin and matrine for 0h,4h,8h and 24 h,q RT-PCR and Western blot were used to evaluate the m RNA and protein expression levels of Omp A of the bacteria in the biofilms on the catheters.Results Compared to those in control group,the expression of Omp A in Escherichia coli were reduced(P<0.05)in all matrine groups and the positive control group in not only middle phase,but also later phase of the biofilm formation.With the reaction time prolonged,the expression of Omp A was further reduced.Yet,there is no significant difference in omp A expression in different concentration matrine group on both protein and m RNA level(P>0.05).Conclusion Matrine can decrease the expression of outer membrane protein A in Escherichia coli biofilm formation in the middle and later phases.Part II The effect of different concentrations of matrine on the biofilm formation in Escherichia coli: the regualtion of Mic AObject To investigate the effect of different concentrations of matrine on the expression of Mic A,a small non-coding RNA of Escherichia coli in the early,middle and later phases of biofilm formation.Methods Biofilms of Escherichia coli(ATCC25922)was induced using several peritoneal dialysis catheters in 24-well plates and divided into 2 tests.One is called restraining test and the catheters were divided into 6 groups: blank control group(negative control),1/2MIC of levofloxacin group(positive control group),1/2MIC,1/4MIC,1/8MIC,and1/16 MIC matrine group.1,3 and 7 days after treatment,the bacterias on the catheters were analyzed.Another called damaging test and the catheters were divided into 5 groups: blank control group(negative control),1MIC,2MIC and 4MIC matrine groups 1MIC of levofloxacin group(positive control group),after 3 days and 7 days culturing,when it is the middle and the later phases of biofilm formation.After treatment of Levofloxacin and matrine for 0h,4h,8h and 24 h,we analysis the E.coli on the catheters.The q RT-PCR were used to evaluate the expression level of Mic A.Results In the restraining test,Compared to those in control group,the expression of Mic A were significantly incresed(P<0.05)in 1/2 MIC and1/4MIC matrine groups in all phases of biofilm formation.And in the damaging test,all matrine groups in not only middle phase,but also later phase of biofilm formation can increase the Mic A level in the E.coli after 4 hours treatment.With the reaction time prolonged,the expression of Mic A tended to be stable.Yet,there is no significant difference in Mic A expression in different concentration of matrine groups(P>0.05).Conclusion Matrine can increase the expression of Mic A in Escherichia coli biofilm formation in different phases of different concentration.Part III The effect of different concentrations of matrine on the biofilm formation in Escherichia coli: the regualtion of ?E.Object To investigate the effect of different concentrations of matrine on the expression of ?E in Escherichia coli in the early,middle and later phases of biofilm formation.Methods Biofilms of Escherichia coli(ATCC25922)was induced using several peritoneal dialysis catheters in 24-well plates and divided into 2 tests.One is called restraining test and the catheters were divided into 6 groups: blank control group(negative control),1/2MIC of levofloxacin group(positive control group),1/2MIC,1/4MIC,1/8MIC,and1/16 MIC matrine group.1,3 and 7 days after treatment,the bacterias on the catheters were analyzed.Another called damaging test and the catheters were divided into 5 groups: blank control group(negative control),1MIC,2MIC and 4MIC matrine groups 1MIC of levofloxacin group(positive control group),after 3 days and 7 days culturing,when it is the middle and the later phases of biofilm formation.After treatment of Levofloxacin and matrine for 0h,4h,8h and 24 h,we analysis the E.coli on the catheters.Western bloting were used to evaluate the expression levels of?E.Results In the restraining test,Compared to those in control group,the expression of ?E were significantly incresed(P<0.05)in all matrine groups in all phases of biofilm formation.And in the damaging test,all matrine groups in not only middle phase,but also later phase of biofilm formation can increase the?E level in the E.coli after 4 hours treatment.With the reaction time prolonged,the expression of ?E tended to be stable or even decline.Yet,there is no significant difference in ?E expression in different concentration of matrine groups(P>0.05).Conclusion Matrine can increase the expression of ?E in Escherichia coli biofilm formation in the early,middle and later phase.