[Objective]To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2( SBD-2) in epithelial cells of ovine oviduct. [Methods]Epithelial cells of ovine oviduct were isolated and cultured; and then the cultured cells at secondary generation were divided into 17β-estradiol( E2,10- 8mol / L) group,estrogen nuclear receptor antagonist ICI182780( 10- 7mol / L) group,PKA antagonist KT-5720( 1 μmol / L) group,PKC antagonist H-7( 50 μmol / L) group,nuclear factor kappa B antagonist PDTC( 50 μmol / L) group and the blank control group( Control). Firstly,different antagonists were added into corresponding antagonist groups in order to interfere the epithelial cells of ovine oviduct for 1 h. Then,17β-estradiol( 10- 8mol / L) was added into each antagonist group and E2 group for cultivation for 6 h. Finally,real-time fluorescent quantitative RT-PCR was used to detect the changes of SBD-2 mRNA expression.[Results]10- 8mol / L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA( P < 0. 05). Estrogen nuclear receptor antagonist ICI182780,NF-κΒ antagonist PDTC and PKC antagonist H-7 could all block the Up-regulatory effects on SBD-2. But PKA antagonist KT-5720 showed no significant effects on the Up-regulation of SBD-2 mRNA expression induced by 17β-estrodiol. [Conclusions] SBD-2 mRNA expression induced by 17β-estrodiol in epithelial cells of ovine oviduct was mediated by estrogen nuclear receptor ICI182780,NF-κB and PKC pathways. However,PKA pathway might not participate in the Up-regulation of SBD-2 mRNA expression.
The paper was to study the effects of different concentrations of lipopolysaccharide(LPS) on expression of nuclear factor Kappa B(NF-κB) and lingual antimicrobial peptide(LAP) gene in mammary epithelial cells of dairy cow.The mammary epithelial cells of dairy cow were stimulated by different concentrations(50,100,200,400 and 800 ng/mL) of LPS.The total RNA of cells was extracted after stimulation for 2,4,8,16,24,48 and 72 h,respectively,and the mRNA expression levels of NF-κB P65 and LAP were evaluated by real-time quantitative PCR.The results showed that the expression of NF-κB P65 and LAP mRNA treated with 400 ng / mL LPS for 72 h were the highest compared to the control group(P<0.01).The result confirmed that the expression activity of NF-κB was enhanced in inflammatory effects of mammary epithelial cells induced by LPS,which regulated the expression of defense gene LAP,with certain dose and time effects.
