脂多糖对奶牛输卵管上皮细胞环氧合酶-2基因表达的调控作用

Regulation Effect of Lipopolysaccharide on Cyclooxygenase-2 Gene Expression in Bovine Oviduct Epithelial Cells

本试验旨在探讨脂多糖(lipopolysaccharide,LPS)对奶牛输卵管上皮细胞环氧合酶-2(cyclooxygenase-2,COX-2)基因表达的影响,阐明LPS对奶牛输卵管上皮细胞COX-2基因表达的调控机制。采用机械法及胰酶消化法分离培养奶牛输卵管上皮细胞;分别将LPS、吲哚美辛、N-乙酰半胱氨酸(N-acetylcysteine,NAC)作用于体外培养的奶牛输卵管上皮细胞,采用实时荧光定量PCR技术检测LPS对奶牛输卵管上皮细胞COX-2基因表达的调控作用。结果显示,不同浓度的LPS能显著促进输卵管上皮细胞COX-2mRNA的表达(P<0.05),且具有浓度依赖性;LPS作用2h时COX-2mRNA的表达量最高,随之表达量逐渐下降到正常水平;NAC能显著抑制LPS诱导的COX-2mRNA表达(P<0.05)。结果表明,LPS能通过NF-κB途径上调奶牛输卵管上皮细胞COX-2mRNA表达量。

The study was aimed to explore the effect of lipopolysaccharide （LPS） on cyclooxygenase-2 （COX-2） expression and explain the regulation mechanism of LPS on COX-2 expression in bovine oviductal epithelial cells. The bovine oviductal epithelial cells were isolated and cultured by using mechanical method and enzyme digestion. Bovine oviduetal epithelial cells were treated by LPS, indomethacin and N-acetylcysteine, respectively. The effect of LPS on the expression of COX-2 mRNA was examined quantitatively by Real-time PCR. The results showed that different concentrations of LPS significantly promoted COX-2 gene expression in bovine epithelial cell （P〈0. 05） in a concentration-dependent manner. After LPS treatment at different time points, it indicated that COX-2 mRNA expression was highest at 2 h after LPS stimulation, and then the expression level decreased gradually to the normal condition; NAC could significantly inhibit COX-2 mRNA expression induced by LPS （P〈0.05）. The expression of COX-2 mRNA in bovine oviductal epithelial cells was increased after LPS stimulation through NF-κB pathway.