Research On Efficacy Of EDTA Combined With Gentamicin And HA In Chronic Bacterial Cystitisof Mice Induced By E.Coli

Part I Preparation of mice models in acute and chronic bacterial cystitisObjectiveTwo different kinds of E.coli strains (ATCC25922and ATCC35218) were separately perfused into the bladders of female Kunming mice by transurethral catheterization. At the second day and14th day we will determine the IBCs and count the numbers of bacteria in the bladder for establishment of acute and chronic bacterial cystitis model in mice.MethodsTwenty four female Kunming mice were randomly into four groups, the bacterial strains (ATCC25922and ATCC35218,1-2x108CFU/ml) were separately inoculated into the bladders by transurethral catheterization. At the second day and14th day, bladders were aseptically harvested and equally divided into two parts. One part was stained with HE and IFA then observe the intracellular bacterial community in the light microscopy and immunofluorescence microscopy. The other halves were homogenized and cultured to count the numbers of colonies.Results 1The IBCs were detected in the superficial mucosa of bladder which prepared by the two kind of strains induced acute cystitis of mice and they can all live in the bladder tissue and the number of colonies werel07CFU/g.2The IBCs were not detected in the superficial mucosa of bladder which prepared by ATCC35218induced chronic cystitis of mice; this E.coli can live persistently in tissue and the number of colonies werel05CFU/g.3The IBCs were not detected in the superficial mucosa of bladder which prepared by ATCC25922induced chronic cystitis of mice; the E.coli can not live persistently in the tissue by the end of two weeks.Conclusion1The bacterial strains ATCC35218and ATCC25922can all induce acute cystitis in female Kunming mice.2The bacterial strains ATCC35218can induce successfully chronic cystitis in female Kunming mice.3The bacterial strains ATCC25922can not induce chronic cystitis in female Kunming mice. Part Ⅱ Induction of urothelial cells shedding by different concentrations of EDTAObjectiveTo investigate the effects of three kinds of EDTA induce urothelial mucosa shedding in the normal mice of female Kunming and select the optimal concentration of EDTA.MethodsMice(n=18)were randomly into three groups and treated with intravesical EDTA solution(30、50、100mmol/L), at the third day mice were killed by cervical dislocation and bladders were aseptically removed. The bladder tissues were stained with HE and IFA, then assess the degree of urothelial damage produced by the EDTA in the light microscopy and immunofluorescence microscopy.Results1Three groups of EDTA can all cause shedding of epithelial cells in the bladder and the damage of Uroplakin.2The injure degree of tissue in the30mmol/L of EDTA were relative less. ConclusionThe30mmol/L of EDTA were the optimal concentration and we will use in the next experiments. Part Ⅲ Treatment effects of EDTA combined with gentamicin and HA on chronic bacterial cystitis of mice induced by E.coliObjectiveTo investigate the treatment effects of EDTA combined with gentamicin and HA on mice of chronic bacterial cystitis induced by E scherichia coli.MethodsMice (n=30) were randomly divided into5groups:EDTA group; EDTA+gentamicin group; EDTA+gentamicin+HA group; negative control group; positive control group. The chronic bacterial cystitis models were established and treated with the above five measures then to analyze the degree of injury about bladder epithelium and calculate the amount of bacteria per gram of tissue.Results1Less injury was observed in the mice treated with EDTA combines gentamicin and HA.2The counts of bacteria living in the bladder tissue were lowest in the group of treated with EDTA combines gentamicin and HA.ConclusionEDTA combine gentamicin and HA can effectively clear off the bacteria in the bladder tissues and it may be a new approach in treatment the chronic UTI.