[ 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^-8 tool/L) group, estrogen nuclear receptor antagonist ICI182780 (10^-7 tool/L) group, PKA antagonist KT-5720 (1 μmol/L) group, PKC antagonist H- 7(50 μmol/L) group, nuclear factor kappa B antagonist PDTC(50μmol/L) group and the blank control group ( Control ). Firstly, different antagonists were added into corresponding antagonist groups in order to interfere the epithelial ceils of ovine oviduct for 1 h. Then, 17β-estradiol ( 10^-8 mol/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^-8 mol/L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA (P 〈 0. 05 ). Estrogen nuclear receptor antagonist ICI182780, NF-κB antagonist PDTC and PKC antagonist H-7 could all block the Up-regnlatory 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 lipopelysaccharide (LPS) on expression of nuclear factor Kappa B (NF-κB) and lingual antimicrobial peptide (LAP) gene in mammary epithelial ceils of dairy cow. The mammary epithelial ceils 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 inammary epithelial cells induced by LPS, which regulated the expression of defense gene LAP, with certain dose and time effects.