Effect Of Polydatin On Chronic Endometritis And NF-?B Pathway In Vitro

Objective: Chronic endometritis produces adverse effectsin patients with assisted reproductive outcome,the treatment is difficult formodern reproductive medicine,clinical routine use of antibiotics for the treatment of chronic endometritis,but the overuse of antibiotics may be the cause of bacterial drug resistance,so this topic through the establishment of human endometrial stromal cells in vitro model of inflammation,giant knotweed glycosides from molecular level research mechanism of anti-inflammatory,provide new ideas for the treatment of chronic endometritis.Method:1)In this study,human endometrial interstitial cells(hESCs)were extracted and cultured by in vitro primary cell culture,and 50% inhibitory concentrations of lipopolysaccharide(LPS)on human endometrial interstitial cells(hESCs)were selected by CCK8 method under different concentrations.2)The selected concentration of LPS was used to induce the inflammation of the endometrial interstitial cells,and the protein expression of the downstream inflammatory cytokines IL-1? and TNF-? induced by LPS was identified by ELISA.3)Through CCK8 selection knotweed nucleoside under the effect of different concentrations on cell proliferation of human endometrial stromal cells,will pass to 2-3 generation of endometrial stromal cells are grown in 96 empty plates after 24 h,to switch to DMEM/F-12 culture medium without serum,at the same time adding different concentrations of polydatin knotweed pretreatment for 24 h,then add half inhibitory concentration of LPS damage cells for 24 h,CCK8 method is used to detect cell vitality.4)Put it will extend the cells grown in 6 empty plate,respectively is: the normal group,LPS group,the giant knotweed glycosides low dose group(25 uM),giant knotweed glycosides in the dose group(50 uM),giant knotweed glycosides(100uM),high dose group after the building given different doses(25uM,50 uM,100uM)of giant knotweed intervention,the normal group and LPS group was given equal without serum DMEM/F-12 culture.Expression levels of proteins associated with NF-?B classic inflammatory pathways(TLR4,IKK?,and p65)were detected byqPT-PCR to observe whether polydatin can reduce the expression level of inflammation-related proteins.Results:1)The median inhibitory concentration of human endometrial mesenchymal cells induced by LPS was 150ug/ml by CCK8 method.2)The levels of IL-1? and TNF-? in the supernatant were significantly higher than those in the control group by ELISA(P<0.05).The inflammatory model was successfully established.3)Polydatin has no toxic effect on human endometrial mesenchymal cell activity in the range of(0-100uM).4)After 12 hours of polydatin pretreatment,CCK8 method was used to detect the cell activity and found that compared with the LPS group,the activity of the endometrial stromal cells in the polydatin pretreatment group was significantly enhanced.5)mRNA expressions of NF-?B inflammatory signaling pathway related proteins TLR4(P< 0.01),IKK(P< 0.01),and p65(P< 0.01)were significantly decreased in the intervention group compared with the LPS group.Conclusion: the protective effect of polydatin is related to the inhibition of NF-?B inflammatory signaling pathway,and plays an active role in the treatment of chronic endometritis,providing a new perspective for the exploration of the pathogenesis of chronic endometritis.