Curcumin For Endometritis Pathogenic Bacteria The Researches On The Antibacterial Activity And The Mechanism Of Anti-inflammatory

Endometritis is a common disease of bovine reproductive system.Endometritis results in the decrease of cattle conception rate,,which seriously affect the development of the cattle industry.The study group found that the principal pathogens causing endometritis in Holstein cows were Staphylococcus aureus and Escherichia coli.The clinical treatment of endometritis using antibiotics and hormone treatment,although the curative effect is the best thing,but it has toxic side effects and affect food safety.Curcumin,an effective component of turmeric,has the effects of anti-inflammation,antioxidation,anti-tumor,lipid-lowering and so on.It has slight toxicity and high safety.In order to introduce curcumin into the application to treat endometritis of cow endometritis,further research on the theoretical basis for the development of anti-inflammatories,improving compatibility and improving curative effect.This study includes the following six aspects:TestⅠTo study the antibacterial effect and mechanism of curcumin for the main pathogenic bacteria of cow endometritis,by Oxford cup method and two times dilution plate method for the determination of curcumin on Bacteriostasis of cow endometritis pathogenic bacteria Staphylococcus aureus,Escherichia coli diameter and minimum inhibitory concentration（MIC）and minimum bactericidal concentration（MBC）,On the basis of above,the antibacterial protection rate of the main pathogenic bacteria infected mice was determined.In order to further explore the antibacterial mechanism of the main pathogenic bacteria of curcumin.The effects of curcumin on the growth curve,cell wall,membrane permeability and membrane potential of the main pathogenic bacteria were determined.The test results show that the protective rate of curcumin and bacteriostasis of mice despite the fact that less than ciprofloxacin,But the test data show that curcumin has bacteriostatic action on main pathogenic bacteria,and it has a good protection for mice which were infected by pathogens.Curcumin has affect for the two kinds of pathogenic bacteria growth curve,cell wall,cell membrane permeability and the membrane potential of the cell membrane.The results showed that Curcumin can have bacteriostasis effect from different aspects,Curcumin could inhibit the cell wall,enhance the permeability of the cell membrane and affect the bioelectricity of cells.TestⅡBecause the pathogenic bacteria caused inflammation in mice and rats,the pathologic response was basically the same,and in order to facilitate the operation,the animals were selected by rats.The effect of curcumin on endometritis and its anti-inflammatory mechanism were further studied in animal experiments.To replicate the model of endometritis in rats with the main pathogenic bacteria in uterine perfusion,Rats were observed before and after building general condition,observe the changes of white blood cell count analysis made classification changes of rat general state,body temperature and blood cell analyzer before and after modeling,compared to the normal group and model group the content of uterus,uterine anatomy of bacterial culture the pathological changes of morphology.Results:Compare to the previous model rats,poor spirit,arch back,loss of appetite,vulva swelling and wet irrigation after the temperature began to rise.Three days after injection in rats,the number of peripheral white blood cells,neutrophils and mononuclear cell percentage increased significantly（P<0.05）,the model of uterine secretions bacterial culture between pathogenic bacteria were seen in the model group compared with normal group,endometrial epithelial cell degeneration and necrosis,edema and congestion,a large number of film the infiltration of inflammatory cells.The results showed that the endometritis model of rat endometritis was successfully replicated by intrauterine infusion pathogenic bacteria.TestⅢIn order to study the effects of curcumin on blood rheology of rats with endometritis.60 female SD rats were randomly divided into 6 groups,normal group,model group,high,medium and low dose curcumin group and ciprofloxacin group,10rats in each group,except the normal group,the other group after drug intervention,7d after medication.With automatic blood rheology analysis of changes of rat blood rheology index of blood analyzer,using enzyme-linked immunosorbent assay（ELISA）detection of serum thromboxane B₂（TXB₂）and 6-to prostaglandin F₁ alpha（6-Keto-PGF₁ alpha）content changes and morphological study on uterus and histopathology.Results:Endometritis rats hemorheology abnormal,after treatment with curcumin,compared with model group,curcumin group rat Whole blood viscosity（WBV）,Plasma viscosity（PV）,Erythrocyte accumulation index（EAI）and Erythrocyte sedimentation rate（ESR）were decreased（P<0.05）;compared with the model group,the content of curcumin in each dose group of 6-Keto-PGF₁ alpha in serum increased significantly（P<0.05）,the content of TXB2 was decreased（P<0.05）.Rat uterine lesions form has improved,the uterus index decreased significantly（P<0.05）,Uterine inflammation pathological changes has different degree of fade,and there are dose dependent.Results:Endometritis with blood stasis syndrome in rats,curcumin can improve blood rheology and microcirculation,promote the regression of endometritis lesions,Its action may increase in serum 6-Ke-to-PGF1 alpha levels,reduce TXB2levels.TestⅣIn order to study the effect of curcumin on the motion of the uterus,30 rats were randomly divided into normal saline group,oxytocin group,curcumin high,medium and low dose groups.The contraction intensity and frequency of the uterus in vitro were observed and recorded before and after administration by BL420E biological signal acquisition system.In order to further explore the mechanism of curcumin on uterus,study the effect of curcumin on rat uterine estrogen endometritis and progesterone on the level,the 60 female SD rats were randomly divided into 6 groups,normal group,model group,oxytocin group,high dose group,curcumin,10 rats in each group,except the normal group,the other group after drug intervention,7d after medication,using radioimmunoassay method to detect the rat uterus tissue E2 and P levels.Results:Compared with normal saline group,Curcumin groups make rats in vitro uterine contraction intensity and frequency were significantly increased（P<0.05）;compared with oxytocin group,of curcumin high,middle dose group of rat uterine contraction intensity and frequency were higher（P<0.05）,curcumin low-dose contraction intensity and oxytocin group there was no significant difference,and the contraction frequency was significantly higher than that of oxytocin group（P<0.05）.compared with the normal group,the level of uterine tissue of E2 rat model group decreased significantly（P<0.05）,P increased significantly（P<0.05）,compared with the model group,the level of E2 in uterine tissue of rats with different doses of curcumin increased significantly（P<0.05）and P were significantly decreased（P<0.05）The results suggest that curcumin can excited uterus in rats,Curcumin can make produce paroxysmal of the uterus to contract,different from the oxytocin tetanic contraction,this contract helps promote inflammatory products in time and speed up the uterine function recovery.curcumin can reduce inflammation increased abnormal uterine estrogen level,and can reduce the inflammation of uterus abnormal increase of progesterone level,which may be the mechanism of curcumin in the treatment of endometritis and excite uterus.TestⅤIn order to further discuss from cell and molecular level of curcumin on experimental rat endometritis curative effect and mechanism of anti-inflammatory,The rats administered with Experiment III,after 7d treatment,the expression of CD14 was detected in endometrial immune group changes,TNF-α,IL-6,IL-8,IL-10 in uterine tissue was detected by ELISA.Results:compared with the model group,Curcumin in each treatment group the expression of CD14 in uterine tissue decreased significantly（P<0.05）;Blood and uterine tissue TNF-a,IL-8,IL-6 content decreased significantly（P<0.05）,the content of IL-10 increased（P<0.05）.Research shows that curcumin is effective in the treatment of endometritis,the mechanism may affect the expression of the CD14,by inhibiting the expression of inflammatory cells on the membrane of CD14,prevented the stimulation of inflammatory mediators on cells,further affect the inflammatory factor TNF-α,IL-6,IL-10,IL-8 secretion,so as to control the occurrence and development of uterine tissue inflammation.TestⅥIn order to further investigate the effects of curcumin on inflammatory signal transduction pathway,experimental animal groups administered as above,the expression of pIKB and IkB,p65 and p-p65,IKKa and TLR4 using Wester-blot method to detect the rat uterine tissue.Wester-blot results showed that compared with the normal group,the expression of model group rats in pIkB,p-p65,TLR4 and IKKa increased（P<0.05）,the expression of Ik B decreased（P<0.05）,indicating bacterial endometritis inflammation and inflammation models TLR4-IKKa-NF-kappa B pathway.The effect of curcumin in uterine tissue of rats under endometritis pI kappa B,p-p65,IKKa and TLR4expression decreased（P<0.05）,increased expression of the IkB（P<0.05）,and the dose dependent.The results suggest that curcumin reduced the expression of TLR4 and down regulation of inflammation in uterine tissue of IKK activation and phosphorylation of Ik B,thereby reducing NF-kB activation,from a variety of ways to prevent TLR4-NF-kB inflammation signal transduction pathway.Thus,inhibiting the release of inflammatory factors such as TNF-α,IL-6,IL-8 and so on,and ultimately take part in the regulation of inflammation and immune response.This study explores the mechanism of curcumin in the treatment of endometritis from different angles,the results show that the effects of curcumin on the main pathogens of cow had good antibacterial effect,good curative effect on rat model of endometritis pathogens infection,proved that curcumin has antibacterial anti-inflammatory and other pharmacological effects,provide a scientific basis for the future on the further development and application of curcumin in clinical treatment of endometritis.