Preliminary Study On The Regulation Of Salidroside By NF-κB And MAPKs Signalling Pathways In Yak Endometritis

Yak breed is the main part of the Qinghai-Tibet plateau grassland animal husbandry.Approximately 14 million heads are existing in that area.Healthy and sustainable breeding of yak plays an important economic role for the whole Tibetan area.Postpartum endometritis seriously affect the reproductive performance of yak.Development of yak industry is facing huge losses due to occurrence of endometritis.Study revealed that E.coli is one of the majorcause of endometritis.E.coli(Escherichia coli).E.coli is one of the main pathogens of cow endometritis.E.coli is the main cause of the infection in uterine epithelial cells,and release the pathogenic bacterial toxins,such as bacterial lipopolysaccharide(LPS).LPS is an important component of gram-negative bacterial cell walls,and is the main factor of gram-negative pathogenic bacteria,it can cause the body to a series of inflammatory response at the moment,still give priority to with antibiotics in clinical treatment of the disease,however,caused by the antibiotics the spread of drug-resistant bacteria and antibiotic residues in food has become a serious problem worldwide,abuse of antibiotics in livestock farming gradually prohibited at the same time,Tibet has always been advocating green farming Therefore,in the prevention and control of yak endometritis,find other treatments to replace antibiotic treatment is imperative in order to find better alternatives,Tibetan medicine has been widely research and application of rhodiola,as a traditional medicine anti-inflammatory adjustment with antioxidant and antibacterial effect in clinical practice can be effective prevention and treatment of endometritis in order to verify whether by large flower rhodiola extract rhodiola glucoside of yak endometrial inflammation have certain control effect,has carried out a series of tests,test contents and results are as follows:1.Yak endometrial cells were cultured in vitro and identified by CK18.The results showed that the cell culture was successful and the purity reached more than 95%.2.With concentration of 1 100 200 500 1000 ng/m L LPS(lipopolysaccharide)bacteria endometrial cells,determined by MTT method is used to detect cell toxicity,calculate its inhibition rate of growth of endometrial cells,in order to determine the optimal concentration of building intervention to concentration of endometrial cells,IL-l beta and production of TNF alpha changes,to see whether the concentration of the inflammatory response results show that the LPS induced in vitro yak endometrial inflammation model of optimal concentration of 100ng/m L,under the concentration has obvious inflammatory process The model of LPS induced inflammation in yak endometrial cells was successfully established3.To preliminary clear rhodiola glucoside in the role of inflammation,experiment directly using escherichia coli for in vitro culture of endometrial cells co-culture,simulate the inflammatory process,on the basis of the control group e.coli group,e.c.with our fabrication:oil+rhodiola glucoside and oil of e.c.with our fabrication:+dexamethasone group,common after the incubation,groups determined by kit SOD activity MDA level,at the same time by ELISA determination of endometrial epithelial cell culture supernatant of TNF alpha IL-1 alpha IL-6 and IL-10 concentration results show that the SOD activity to reduce the inflammation induced by E.coli group MDA levels rhodiola glycosides and dexamethasone pretreatment group of endometrial epithelial cells can increase SOD activity decrease MDA level treatment group can significantly increase the e.coli cells inflammatory factor TNF alpha IL-1 alpha,the production of IL-6 and IL-10,rhodiola glycosides and dexamethasone treatment group can significantly reduce the level of inflammatory factors,rhodiola glucoside of endometritis has certain anti-inflammatory effect.4.In order to preliminarily study the role of salidroside in regulating yak endometritis through NF-B and MAPKs signaling pathways,the LPS control group,LPS+DEX group,LPS+salidroside group,and IHC were used to detect the MPO activity at different treatment levels after 24 h incubation.The expression levels of NF-B TNF-il-1 il-6 il-10 Mc P-1m RNA were detected by ELISA and q PCR.Western Blot detection of MPO activity of signaling pathway related protein I B p65 p38 EPK JNK showed that salidroside(20,40 and 80 ng/m L)could slightly improve the cell viability of yak uterine epithelial cells,but the difference was not significant(P>0.05)ELISA and q PCR showed that the expression levels of related factors in the LPS group were increased,and salidroside significantly inhibited the expression of these factors(P<0.05),especially the concentration of 80 ng/m L,rhodiola rosea is most close to the dexamethasone group and control group in Western Blot test results show that rhodiola glycosides and dexamethasone in the treatment group related protein expression levels drop,rhodiola glycosides show can significantly inhibit the related protein expression of ascension that rhodiola glucoside can through the nf-kappa B and MAPKs signaling pathways,lower levels of inflammatory factors,treatment of yak endometrial injury effect is close to the effect of dexamethasone,can consider to be used in the treatment of yak endometrial lesions This study provides a reference for the future prevention and treatment of endometritis in yaks.