| The study was aimed to identify the main pathogens from cow,Escherichia coli and Staphylococcus aureus,and to study the mechanism Mongolian Medicine Danggui-4 Wei San of medicine inhibiting pathogenic Escherichia coli and Staphylococcus aureus from Daily cow endometritis antibacterial mechanism and effect on the mechanism of endometritis in rats infected with pathogenic E.coli and S.aureus the mechanism of action,and suffer from endometriosis induced by colitis in rats to make a detailed study.This study not only lay a foundation for the development of Chinese herbal medicine and probiotics but also to provide data and theoretical basis for clinical application in the future.This study included four experiments.The purpose of the experiment 1:Purulent or mucopurulent uterine exudates in the vagina occurring after the third week of lactation from 65 postpartum cows were collected by using uterine swabs.Rat’s uterus test,PCR,and susceptibility paper method were used to detect pathogenic.Here,we report evidence that:Rat’s uterus test,PCR,and susceptibility paper method were used to detect pathogenicity.The results showed that this rat’s uterine perfused with mixed bacteria(1:1,1.0×109 cfu/ml)displayed uterine congestion,swelling,thickening of the uterine internal pyometra phenomenons after 24 hours.The virulence genes of yjaA,KpsMII,hlyA,sfas and irp2 were detected for in the 27 pathogenic E.coli isolates and virulence genes ChuA and PapC were negative,and the virulence genes of TSST-1,ermA,ermC,Blaz and SEB were detected for in the 32 pathogenic S.aureus isolates and virulence genes etb,SEJ and SEE were negative.In randomly selected ten sick cows and most isolated virulence genes isolates were tolerance to penicillin,doxycycline,polymyxin B,amoxicillin,tetracycline,and other antibiotics.The purpose of experiment 2 was designed on the basis of the experiment 2,Using ELISA turbidimetric method,kit method and o-nitrobenzoic beta-D-synthesis(ONPG)method of Mongolian Medicine Danggui-4 Wei San and nisin on E.coli and S.aureus bacteria growth curve,membrane potential,the content of cell wall and cell membrane permeability.①The antibacterial Mongolian Medicine Danggui-4 Wei San and nisin medicine the growth of pathogenic S.aureus,however,nisin has barely bacteriostatic effect of pathogenic E.coli.The experiment 3 was to study its pathogenic.Rat’s uterus test,PCR,and susceptibility paper method were used to detect pathogenicity.The results showed that this rat’s uterine perfused with mixed bacteria(1:1,1.0×109 cfu/ml)displayed uterine congestion,swelling,thickening of the uterine internal pyometra phenomenons after 72 hours.The experiment 3 was selected seventy SD rats divided into seven groups,nisin(0.1 mg/mL),the model group,saline solution(negative control group,20 g/kg·b·w),ciprofloxacin(0.13 g/kg;positive control),the high dose of antibacterial compound rhubarb(1 g/kg(b·w)),the medium dose of antibacterial Mongolian Medicine Danggui-4 Wei San(0.5 g/kg(b·w)and the low dose of Mongolian Medicine Danggui-4 Wei San(0.025 g/kg(b·w)),respectively.At the end of the experiment,the samples of blood and tissues was collected for detection of relevant indicators of terus.Thereby,the experiment 5 was conducted to study antibacterial Mongolian Medicine Danggui-4 Wei San and nisin medicine effects on the organ-organ mechanism of action,and in rats infected with pathogenic E.coli and S.aureus,and the mechanism of action,and the samples of tissues was collected for detection of relevant indicators of colonic.①Compared with negative control group(P<0.05),the concentration of 17β-Estradiol,P4 and oxytocin in blood were significantly improved in the middle and high dose of the antibacterial Mongolian Medicine Danggui-4 Wei San group and the positive control group,but model group was gradually reduced in the positive control group.However,The factor,IL-6,IL-10,CINC-1,TNF-α and COX-2,were significantly increase in the model group compared with other drug group,and the levels,IL-6,IL-10,CINC-1,TNF-α and COX-2 in serum on endometrial rats were significantly lower compared with model group and the negative control group(P<0.05).④From 0 h to 72 h,the concentration of MMP-9 and TIMP-1 in the model group was significantly lower than those in the normal saline group(P<0.05)and the concentration of MMP-2 and TIMP-2 decreases gradually.In the nisin and high dose of antibacterial Mongolian Medicine Danggui-4 Wei San group,the concentration of MMP-9,TIMP-1,MMP-2 and TIMP-2 were significantly higher than those in the normal saline group(P<0.05),in which the nisin group and high dose ofantibacterial Mongolian Medicine Danggui-4 Wei San group showed the best effects.Experiment 4:①The concentration of MPO,TBX2 and oxytocin(OXT)in the uterine tissue homogenate of rats with endometritis was significantly improved than that in the model group(P<0.05)after 72 h of modeling,especially the concentration of oxytocin,TBX2 and MPO in nisin group and high dose group of the antibacterial Mongolian Medicine Danggui-4 Wei San group was significantly improved,which indicated that the concentration of 6-K-PGF1α was significantly improved and the ratio of PGI2/TXA2 was maintained Hemostasis does not leave stasis,②In colonic tissue,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 were tested by ELISA.The contents of TNF-α,IL-1β,IL-6 and IL-8 were significantly decreased in the positive control group and high dose group of the antibacterial Mongolian Medicine Danggui-4 Wei San group compared with the negative control group(P<0.05).③Compared with negative control group and the model group,the concentration of IL-2 and IFN-y in the high dose group of the antibacterial Mongolian Medicine Danggui-4 Wei San group and the positive control group was improved(P<0.05).In conclusion,in serum on endometrial rats,the contents of 17β-Estradiol,P4,OXT,IL-6,IL-10,CINC-1,TNF-α,COX-2,MMP-9/TIMP-1 and MMP-2/TIMP-2 in the the middle and high dose of the antibacterial Mongolian Medicine Danggui-4 Wei San group and nisin group,as well as in its uterine tissue,the contents of MPO,OXT,TXB2 and 6-K-PGF1α,and the contents of TNF-α,IL-1β,IL-6,IL-8,IL-10,IL-2 and IFN-γ in its colonic tissue were all positively regulated. |
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