Effect Of β-Hydroxybutyrate On Synthesis And Secretion Of Prostaglandin E2 In Bovine Endometrial Cells

Background:Dairy cow ketosis is a metabolic disease that occurs frequently in perinatal dairy cows.Β-hydroxybutyric acid(BHBA)is the main component of ketone bodies.High levels of BHBA can induce oxidative stress and inflammatory reactions in dairy cows,resulting in decreased lactation and postpartum disease,which seriously affects the reproductive performance of dairy cows.Prostaglandins are a kind of physiologically active unsaturated fatty acids,which are widely distributed in various tissues and body fluids of the body,and are closely related to various life activities such as inflammation and reproduction.OBJECTIVE:The purpose of this study was to investigate the changes of prostaglandin E2 and the expression of genes involved in the synthesis of prostaglandin E2 in BEND cells treated with BHBA in vitro by BNE,and to elucidate the BHBA-to-BEND cell prostaglandin E2.The effects of synthesis and secretion provide a theoretical basis for further study on the effects of ketosis on postpartum production and reproductive performance of dairy cows and their mechanisms.METHODS:Four different concentrations of BHBA were added to cultured bovine endometrial cells(BEND)in vitro,and cells were extracted at 6 h,12 h,and 24 h.The supernatant,total RNA and total protein were cultured,the content of prostaglandin E2 was detected by ELISA,and the expression levels of prostatic synthesis pathway-related genes such as cPLA2,COX-2 and cPGEs were detected by real-time PCR and Western-blot.Variety.RESULTS:ELISA results showed that BEND cells treated with different concentrations of BHBA could significantly reduce the content of PGE2 in BEND cell culture supernatants at 12 h and 24 h,but the effect was not significant at 6 h.The results of fluorescence quantitative PCR showed that different concentrations of BHBA were treated.After 6 h of BEND cells,the expressions of cPLA2,COX-2 and cPGEs mRNA did not change significantly.However,after 12 h and 24 h,BHBA could significantly promote the expression of cPLA2 mRNA and inhibit the expression of COX-2 and cPGEs mRNA in time and dose.Dependence;WB results showed that BEND cells treated with different concentrations of BHBA for 12 h significantly promoted cPLA2 protein expression and inhibited COX-2 and cPGEs protein expression.Conclusion:BHBA can significantly reduce the content of PGE2 in BEND cell supernatant,which may be achieved by regulating the changes of key gene expression levels of PGE2 synthesis pathways such as cPLA2,COX-2 and cPGEs in BEND cells.BHBA can occur directly after 12 hours of cell action.