| Urinary tract infection refers to inflammation of the urinary tract, which caused by pathogens reproduce in the urinary tract of organism and encroach on mucosa or tissue of urinary tract. It is one of clinically the most common infectious diseases. It caused by bacterial infection and E. coli is the most common. While E. coli caused by urinary tract infections also known as pathogenic E. coli or Uropathogenic E. coli. In the early stages of this study, we had also conducted a series of studies on the pathogenesis and treatment methods of the UPEC bacteria urinary tract infection, and found that the key and difficulty of clinical therapy is bacterial resistance. In order to achieve the desired results, we further looking for a safe and effective treatment program of antibiotics to substitute has a very important practical significance, and which is also to play to the advantages of Traditional Chinese Medicine (TCM)syndrome differentiation. Therefore, the search of the new and therapeutic value compound traditional has become an important content about the urinary tract infection study. Therefore, the main task and rule of TCM treatment of urinary tract infections is heat and dampness. Traditional Chinese Medicine of Fezhengqingrelishi, which contains more than ten kinds of herbal medicines, from the perspective of TCM syndrome differentiation treatment, it can be reached heat and dampness tonglin, Yin and kidney and spleen to rousing the therapeutic effect.Therefore, in this paper, UPEC10screened out through the preliminary results as the research object, which has high pathogenicity, strong drug resistance, biofilm-positive and ESBL-positive.And by the urethral Intubation to establish the UPEC urinary tract infection in a mouse model, to interfere the mouse model by Traditional Chinese Medicine of Fezhengqingrelishi and antibiotics (imipenem), and by monitoring Mice of symptoms, signs, and various physiological changes before and after treatment to assess the therapeutic effect of righting the heat and dampness compound recipe to UPEC bacteria urinary tract infection. Finally, we conclude that Traditional Chinese Medicine of Fezhengqingrelishi has a therapeutic effect on multiple drug-resistant UPEC bacteria urinary tract infection, and be quite with antibiotic imipenem.The main subject of study carry out from the following aspects:First of all,24virulence genes widely distribute of95UPEC clinical isolates were selected, were obtained15UPEC bacteria of strong pathogenic according to the early study of our laboratory on the chinese herbal drugs antibiosis and the analyzed results of E. coli virulence factors as well as the distribution and role of the resistance gene which have been reported at home and abroad; By analyzing the drug resistance spectrum and correlation detection of virulence phenotype of15UPEC bacteria of strong pathogenic, we found that UPEC5, UPEC10, UPEC169strains showed hemolysis, of which UPEC10and UPEC169with high rate of the common antibiotics belong to resistance biofilm-positive strains, simultaneously, UPEC10strain of ESBL-positive with the higher drug tolerance.And then we combined the common antibiotic resistance by95UPEC clinical isolates to analysis the internal relationship between the resistance and the distribution of the virulence and resistance genes of the isolates. As a result, the number of virulence and resistance genes significantly related to the degree of antibiotic resistance, the more Virulence genes isolates with, the stronger drug tolerance, UPEC strains of ESBL-positive with the high distribution rate of resistance genes, as well as the common antibiotics resistance.Then, in the process of constructing a green fluorescence of GFP marker strain, the key is to screen the UPEC strain for urinary tract infection model. According to preliminary results, we chose the UPEC10as the strain of this experimental study which has high pathogenicity, strong drug resistance, biofilm-positive and ESBL-positive. We adopted the electrotransfer method to transfer GFP fluorescence plasmid to the experimental strain, and observed fluorescence bacterial under the fluorescence microscope, ultimately, we found that GFP fluorescence plasmid had been transferred to UPEC10and the fluorescently labeled UPEC10-GFP strain was constructed successfully. That has laid a certain foundation for late stage in determining the model of urinary tract infection successfully constructed and grasping the differences of the dynamic changes in vivo in urinary tract infections process.Through the preliminary study on the antimicrobial mechanism, we adopted the microdilution method to test the MIC/MBC as well drug resistance of TCM. We found that the minimum inhibitory concentration (MIC) of TCM on UPEC10bacteria is125mg/ml and the minimum bactericidal concentration MBC is250mg/ml; simultaneously inverse to the drug resistance of UPEC10bacteria. At the same time, we adopted the MTT and microdilution method to test the formation of biofilm as well the effect on the formation of biofilm of TCM. We conclude that TCM has a certain inhibitory effect on biofilm formation whether in the pre-coated or not; while the presence of biofilm has enhanced antibiotics resistance of UPEC10.Finally, in the animal experiments, through exploring and testing, we obtained the optimal infected concentration and dose of UPEC10-GFP bacteria, and established a infecting mouse model. Then by Traditional Chinese Medicine of Fezhengqingrelishi and antibiotics (imipenem) to interfere in the mouse model. Finally, by colony counting spleen, kidney, bladder, and urine of mice and analyzing bacterial clearance rate of organs to assess the therapeutic effect of herbal compound recipe to multiple drug-resistant UPEC10-GFP bacteria urinary tract infection. As a result, by the therapeutic of Traditional Chinese Medicine of Fezhengqingrelishi, bacterial counts in various organs were lower than the model group and the antibiotic group, and the urine bacteria count as well as organ index were lower than the model group, but quite the antibiotic group. Simultaneously, the containing serum of prepared by intragastric administration on mice has a certain inhibitory effect on bacteria.Through the above experiments, we can be further concluded that the presence of biofilm reduced the drug resistance of UPEC bacteria and so even more detrimental to the growth of bacteria, while TCM can inhibit the biofilm formation of UPEC and reduce the colonization and survival ability of UPEC bacteria in mice, therefore, the capacity of TCM inhibit UPEC bacteria parasitized in Organs may be superior to antibiotics during the treatment. By the therapeutics of TCM, although the bacteria still partially exists, but can inhibit the bacteria parasitic in the organs, simultaneously, organ inflammatory injury has also correspondingly reduced through the biofilm change the appearance.In summary, traditional Chinese Medicine of Fezhengqingrelishi has a therapeutic effect on multiple drug-resistant UPEC bacteria urinary tract infection, and be quite with antibiotic imipenem. But, TCM and antibiotics are not able to remove the UPEC bacterias completely from the mice in vivo, which may have a certain relationship with UPEC bacteria can proliferate by itself or UPEC induced relapse. By the therapeutics of TCM, the biofilm changed the appearance can reduce the organ inflammatory injury.Therefore, Traditional Chinese Medicine of Fezhengqingrelishi has an important practical significance in treating urinary tract infections which the UPEC bacteria caused. However, To a certain extent, TCM of Fezhengqingrelishi can reduce the drug resistance of bacteria and increase the sensitivity to antibiotics, which prompts that it may be has a certain relationship with some plasmid eliminating pathogens; And so the concrete and effective mechanisms of its reversal bacterial resistance can be further studied. |
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