| This paper mainly studies the extraction of Zanthoxylum Bungeanum essential oil,chemical composition analysis and antibacterial activity,optimum extraction conditions were determined by single factor test and response surface design method;and the main chemical components of Zanthoxylum Bungeanum essential oil were determined by GC-MS analysis method.Inhibition by in vivo and in vitro antibacterial experiment of Zanthoxylum Bungeanum essential oil on Escherichia coli,and research the in vitro antibacterial mechanism,as well as from the molecular level study of Zanthoxylum Bungeanum essential oil induced inhibition of intestinal infection of Escherichia coli.The main research contents and results are as follows:1?The extraction of Zanthoxylum Bungeanum essential oil.Supercritical carbon dioxide extraction was used to extract the Zanthoxylum bungeanum essential oil.The optimum extraction conditions were obtained by single factor test and response surface analysis.Conditions: extraction pressure 26.3Mpa,extraction time 2.53 h,extraction temperature 50?,extraction CO2 flow 21L/h,extraction the mesh 60.The Zanthoxylum Bungeanum essential oil yield is 11.15%.2?Analysis of the main chemical components of Zanthoxylum Bungeanum essential oil.By gas chromatography-mass spectrometry GC-MS analysis method on the main chemical components of essential oil were analyzed,the analysis results obtained Zanthoxylum Bungeanum essential oil mainly contain two kinds of compounds are hydrocarbon oxygenated organic compounds and terpene compounds.Among which 24 kinds of alkenes,accounted for 41.372%;12 kinds of alcohol compounds,accounted for 18.923%;13 kinds of ester compounds,accounting for35.870%;5 kinds of ketone compounds,accounting for 2.330%;2 kinds of aldehydes,accounted for 0.157%;3 kinds of acid compounds,accounted for 1.348%.The high content of several compounds of ZBEO were 2,6,9,11-Dodecatetraene-1-Carboxylic acid methyl ester?12.428%?,4-Terpinenylacetate?11.249%?,Caryophyllene oxide?8.768%?,D-Limonene?7.902%?,Eucalyptol?5.935%?,?-Terpineol acetate?4.533%?.According to the report,ZBEO had the function of inhibiting bacteria,and these main components had played a very important role.3?Study on activity and mechanism of Zanthoxylum Bungeanum essential oil in vitro.The inhibitory effect of in vitro antibacterial experiment of ZanthoxylumBungeanum essential oil on Escherichia coli.By two times dilution method,get the minimum inhibitory concentration of Zanthoxylum Bungeanum essential oil inhibiting Escherichia coli 24 mg.mL-1,and bacterial growth in extended period and add pepper essential oil on the number of early,bacterial growth was significantly suppression;filter paper method was used to get the the antibacterial circle of Zanthoxylum Bungeanum essential oil diameter is 20mmą1.0mm;results show that Zanthoxylum Bungeanum essential oil can inhibit Escherichia coli.The effect of strong staining by gram,SEM observation and bacterial cell dissolution determination,preliminary determination of the antibacterial mechanism of Zanthoxylum Bungeanum essential oil is to destroy the structure of Escherichia coli the integrity of the cell,leading to cell membrane rupture,cell content,resulting in the death of bacteria.4?Study on the activity and molecular mechanism of antibacterial activity of intestinal infection of Zanthoxylum Bungeanum essential oil caused by Escherichia coli.By intraperitoneal injection of Escherichia coli in mice to establish a mouse model of diarrhea,antibacterial activity of essential oil in vivo.The experimental mice were divided into 6 groups: normal control group,model group,positive control group?berberine?,pepper oil high dose group,dose group of Zanthoxylum Bungeanum essential oil,low dose group,administered continuously for 6 days.The mice symptom score,diarrhea rate,and body weight,food intake and water intake statistical analysis results showed that the essential oil has a certain therapeutic effect on mice with diarrhea,and the higher the concentration,the better the treatment effect.According to the pathological observation of small intestine of mice the results show that the intestinal tissue of model group was destroyed,atrophy of the intestinal villi broken off significantly,the fuzzy structure of intestinal gland tissue,submucosal edema,infiltration of inflammatory cells,and epithelial tissue structure.High dose group curative effect is very obvious,the intestinal villi were orderly,rarely swelling off,no inflammatory cell infiltration,intestinal gland hyperplasia,submucosal edema,epithelial tissue integrity.Low dose group intestinal structure improvement effect is not obvious,villi broken off,intestinal gland hyperplasia is not prominent no,epithelial tissue integrity,but compared with the model group,or to a certain extent.So that Zanthoxylum Bungeanum essential oil has obvious improvement effect on Escherichia coli induced diarrhea in mice,and the higher the concentration effect is more obvious.In the pathogenesis of Escherichia coli induced diarrhea in a variety of inflammatory cytokines play an important role.This paper studies the Toll like receptor2?TLR-2?in small intestine of mice,Toll like receptor 4?TLR-4?,tumor necrosis factor alpha?TNF-??,interleukin 6?IL-6?,white blood cell interleukin 8?IL-8?expression of inflammatory cytokine genes,and the intervention effect of Zanthoxylum Bungeanum essential oil.The results showed that TLR-2,intestinal infection in mice induced by Escherichia coli in small intestine of TLR-4,TNF-?,IL-6,IL-8 gene expression was significantly increased by Zanthoxylum Bungeanum essential oil after intervention,TLR-2,TLR-4 in small intestine TNF-?,IL-6,IL-8 gene expression was significantly decreased,indicating the mechanism of Zanthoxylum Bungeanum essential oil inhibit Escherichia coli infection caused by intestinal dysfunction may be associated with regulation of inflammatory cytokine expression.In summary,using supercritical carbon dioxide extraction method can get the better of the Zanthoxylum Bungeanum essential oil extraction process,and the main chemical components of the Zanthoxylum Bungeanum essential oil were analyzed.Zanthoxylum Bungeanum essential oil has inhibitory effect on Escherichia coli in vitro and in vivo significantly,has obvious effect on Escherichia coli induced diarrhea and intestinal tissue of mice loss. |
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