The Inhibition Effect Of Agastache Rugosa And Origanum Vulgare L. On Drug Resistance Of ESBLs-producing E. Coli

Objectives:To definite the drug resistance level and preliminarily form screening scheme of clinical medication for ESBLs-producing E.coli,which are based on the study of distribution situation,drug resistance,ESBLs genotypes of ESBLs-producing E.coli,collected from human.On the basis of prevention and control,to explore action of extracts from Agastache rugosa and Origanum vulgare L.for ESBLs-producing E.coli,which practical combine clinic drug resistant problem with screening of drug resistant inhibitors from traditional Chinese medicine.It is designed to interdict or reduce the produce and spread of ESBLs-producing E.coli,which offer research basis for prevention and treatment of clinic drug-resistance E.coli.Methods:? With the method of microbiology and epidemiology to study distribution situation and drug resistance of ESBLs-producing E.coli from some hospitals in Urumqi city.? Applied molecular biology,molecular epidemiology,Polymerase Chain Reaction technology and so on to explore the characteristic of ESBLs genotypes from ESBLs-producing E.coli.? Analysed chemical components and determined contents of extracts from Agastache rugosa and Origanum vulgare L.by Gas Chromatography-Mass Spectrometer method and some chemical method of traditional Chinese medicine.? Utilized principal component analysis and clustering analysis to study differences among essential oils from six different production areas of Origanum vulgare L.and optimize better resource.? Determined antibacterial activities of extracts from Agastache rugosa and Origanum vulgare L.to Staphylococcus aureus,E.coli and ESBLs-producing E.coli with the methods of broth dilution method and disc diffusion method(modified Kirby-Bauer method).? Utilized PCR method and 96-hole method to study inhibition effect of antibacterial active single component from Agastache rugosa and Origanum vulgare L.on ESBLs gene and biofilm formation of ESBLs-producing E.coli at the molecular level and the overall level.Results:? The specimen types mainly distribute in urine,sputum,blood,appendix content specimen,and higher infection rates in departments of general surgery,respiratory,neurological surgery,urology,kidney disease.? Statistic analysis of drug-resistant spectrum that ESBLs-producing E.coli has higher resistance rate for ?-lactam antibiotics and imipenem,meropenem of carbapenems antibiotics,which sensitive rates are 100%so far.? The main ESBLs genotype of ESBLs-producing E.coli is CTX-M-14 type,which is relative conservative,mutation site is few and mutation risk is lower.? With the method of GC-MS,the 21 kinds chemical components of essential oil from Agastache rugosa have been identified,representing 99.78%of the total oil.The major compounds were pulegone(34.10%),estragole(29.54%),p-Menthan-3-one(1R,4R)-(+)(16.70%),limonene(8.18%).According to classification of chemical components,the oxygenated monoterpenes,monoterpenes and sesquiterpenoids of essential oil from Agastache rugosa account for 55.29%,8.69%,3.56%,respectively.The content of total flavonoids from Agastache rugosa is 28.52 mg/g.? This procedure allowed the identification of 11 to 46 components among six production areas of Origanum vulgare L.,representing 98.5%to 99.9%of the total oil extracted.The common components are ?-citronellol,citronellol acetate,thymol,aryophyllene.According to classification of chemical components,monoterpenes,oxygenated monoterpenes,sesquiterpenoids and oxygenated sesquiterpenoids exist in different essential oils.The oxygenated monoterpenes was predominant in all the essential oils.? From the principal component analysis and cluster analysis of chemical components of essential oils from six different production areas of Origanum vulgare L.,which obtained three principal components,the first main component are eucalyptol and eugenol methyl ether;the second major component are germacrene D and aryophyllene oxide;and the third major component is thymol.The cumulative contribution rates of the three main components reach 99.57%.According to the principal component factor and weighted comprehensive scores,Shanqiu of Henan,Anhui and Yili have better quality.? The essential oils from Agastache rugosa and Origanum vulgare L.have inhibition and bactericidal action to Staphylococcus aureus and E.coli.Meanwhile,the total flavonoids from Agastache rugosa have poor inhibition effect.?Screened out active components,including pulegone,?-citronellol,eucalyptol and thymol,as the single active components,Pulegone,?-citronellol,eucalyptol have better inhibition effect,except thymol.? The single active components have little influence on the sequence of ESBLs gene.However,it inhibited the biofilm formation of ESBLs-producing E.coli to influence the bacteria growth.Conclusions:The ESBLs-producing E.coli has a wide distribution and resistance rate rise year by year,which isolated from some hospitals in Urumqi city.The main ESBLs genotype of ESBLs-producing E.coli is CTX-M-14 type,screened out active components,including Pulegone,?-citronellol,eucalyptol have little influence on the sequence of ESBLs gene.However,it inhibited the biofilm formation of ESBLs-producing E.coli to influence the bacteria growth.It is need to further research and explore whether change the related regulation mechanism of biofilm,and provide the research basis for further explore and study the resistant inhibit drugs with high efficiency and low toxicity.