Study On The Prescription Of Traditional Chinese Medicine Based On The Mechanism Of E.coli β-lactam Drug Resistance Elimination

With the emergence of multi-drug resistance of E.coli,the therapeutic effect of antibiotics has become worse and worse.Traditional Chinese medicine has the effect of multi-target elimination of E.coli resistance due to its complex composition.At present,researches on the elimination of E.coli resistance with traditional Chinese medicine are based on the effect of elimination of E.coli resistance in vitro,and there is no clear prescription screening method based on the mechanism of elimination of E.coli resistance.Therefore,firstly select single Chinese medicines with better anti-drug effects according to different resistance mechanisms,and then formulate prescriptions based on the principle of omni-directional and multi-target anti-drug elimination;secondly,they are preliminarily screened out through in vitro anti-drug elimination effect and FIC test It is a traditional Chinese medicine prescription with additive or synergistic effect with cefotaxime.Finally,the E.coli animal model is used to determine the best Chinese medicine prescriptions for the elimination of drug resistance in clinical trials,and provide scientific research basis for clinical screening of new inhibitors.1.Screening of traditional Chinese medicine based onβ-lactam drug resistance mechanism of chicken-derived E.coli.Four strains of multidrug-resistant Escherichia coli were co-cultured with traditional Chinese medicine and four strains of bacteria for 72 h.Then the bacterial R plasmid,the activity of extended-spectrumβ-lactamases（ESBLs）,the relative expression of active efflux genes Acr A-m RNA,Acr B-m RNA and the change of biofilm adhesion were detected by gel electrophoresis,double-disk method,fluorescence quantitative PCR and biofilm positive screening method.The results showed that Coptis chinensis had good inhibitory effect on ESBLs activity of resistant Escherichia coli.Rhizoma Coptidis,Herba Tongue and Herba Euphorbiae have good inhibitory effect on active efflux system of resistant Escherichia coli.Rhizoma Coptidis,Herba Tongue,dandelion and Herba Euphorbiae have good effect on reducing the adhesion of drug-resistant Escherichia coli biofilm.2.Study on traditional Chinese medicine prescription based on the elimination effect of drug resistance of chicken Escherichia coli in vitro.According to the previous research basis and the results of this study,hawthorn+Coptis chinensis and hawthorn+Tonguecao were selected according to different proportions,and then Escherichia coli O₇₈（resistant to cefotaxime）was used as the experimental object.Oxford cup method was used to determine the in vitro antibacterial effect of the prescription+cefotaxime,and the compatibility ratio of the prescription with good in vitro drug resistance elimination effect was screened.Finally,the synergistic effect of the prescription and cefotaxime was evaluated by FIC method.The results showed that hawthorn+hyoid（2:1）+cefotaxime and hawthorn+Coptis chinensis（1:1）+cefotaxime had the best elimination effect on E.coli O₇₈in vitro,and FIC was judged as additive effect and synergistic effect.3.Based on animal infection model clinical treatment effect of traditional Chinese medicine prescription research.The infection model of laying hens was established by injecting E.coli O₇₈into abdominal air sac.The sick chickens were treated with hawthorn+tongue grass（2:1）+cefotaxime and hawthorn+Coptis chinensis（1:1）+cefotaxime.The results showed that the cure rate was 90%,the effective rate was 93.3%,and the mortality rate was 6.7%in group 1,hawthorn+Coptis（1:1）+cefotaxime treatment.Test group 2,namely hawthorn+tongue grass（2:1）+cefotaxime treatment of chickens,the cure rate was 80%,effective rate was83.3%,mortality was 13.3%.In summary,the combination of hawthorn+Coptis chinensis（1:1）and hawthorn+Tonguecao（2:1）,which were composed of the elimination mechanism of traditional Chinese medicine forβ-lactam drug resistance of E.coli,had better in vitro drug resistance elimination effect,and the animal clinical trials showed that hawthorn+Coptis chinensis（1:1）had better therapeutic effect on resistant E.coli disease.