前列腺素E2和F2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响

doi:10.16431/j.cnki.1671-7236.2017.09.025

摘要/Abstract

摘要：

试验旨在阐明前列腺素E₂（prostaglandin E₂，PGE₂）和F_2α（prostaglandin F_2α，PGF_2α）对体外培养的奶牛子宫内膜上皮细胞中环氧合酶-1（cyclooxygenase-1，COX-1））与环氧合酶-2（cyclooxygenase-2，COX-2）表达的影响。培养奶牛子宫内膜上皮原代细胞和传代细胞，第4代细胞以1×10⁶个/孔接种于6孔板，以10^-7mol/L PGE₂和PGF_2α分别预处理细胞24 h，以100 ng/mL细菌脂多糖（lipopolysaccharides，LPS）刺激细胞4、8和12 h后分别提取RNA和总蛋白质，采用实时荧光定量PCR与Western blotting等技术检测COX-1与COX-2 mRNA和蛋白质的表达量。结果表明，与对照组相比，COX-1 mRNA表达量在PGE₂单独作用4、8和12 h后显著上调（P<0.05）；COX-2 mRNA表达量在PGE₂单独作用4和12 h后显著上调（P<0.05），PGE₂单独处理使COX-1、COX-2蛋白表达量均显著上调（P<0.05）。与对照组相比，LPS刺激8和12 h时COX-1 mRNA表达量显著下调（P<0.05），LPS刺激后COX-1蛋白表达量无显著变化（P>0.05）；LPS刺激后4、8和12 h时COX-2 mRNA表达量显著上调（P<0.05），LPS刺激后COX-2蛋白表达量显著上调（P<0.05）。与LPS单独处理组相比，LPS+PGE₂处理组在8和12 h时COX-1和COX-2 mRNA表达量均显著上调（P<0.05），同时COX-1和COX-2蛋白表达量也显著上调（P<0.05）。PGF_2α在LPS未刺激和刺激后对COX-1和COX-2 mRNA的表达无显著影响（P>0.05），仅在PGF_2α单独处理8和12 h后COX-1 mRNA表达量上调（P<0.05）。两种激素联合处理与各自单独处理及LPS单独刺激相比，对COX-1和COX-2 mRNA表达具有一定的协同诱导作用。

关键词: 奶牛子宫内膜上皮细胞; 环氧合酶-1; 环氧合酶-2; 前列腺素; 脂多糖

Abstract:

This study was aimed to clarify the effects of prostaglandin E₂ (PGE₂) and prostaglandin F_2α (PGF_2α) on cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) mRNA and protein expression in bovine endometrial epithelial cells. Primary and generation of bovine endometrial epithelial cells were cultured, and the fourth generation cells with 1×10⁶/hole vaccination were pretreated with PGE₂, PGF_2α, PGE₂+PGF_2α (10^-7 mol/L) for 24 h, respectively, following treatment with 100 ng/mL LPS for 4, 8 and 12 h, respectively. The expression of COX-1 and COX-2 mRNA and protein were measured by Real-time quantitative PCR and Western blotting methods. The results showed that compare with control group, the expression of COX-1 mRNA was significantly up-regulated after PGE₂ treatment at 4, 8 and 12 h, the expression of COX-2 mRNA was significantly up-regulated at 4 and 12 h, the COX-1 and COX-2 proteins were significantly up-regulated after PGE₂ treatment (P<0.05). Compare with control group, the expression of COX-1 mRNA was significantly dowm-regulated by LPS treatment at 8 and 12 h, while the COX-1 protein was not affected, the expression of COX-2 mRNA was significantly up-regulated by LPS treatment at 4, 8 and 12 h,and COX-2 protein was significantly up-regulated after LPS treatment (P<0.05). Compared with LPS treatment group, the expression of COX-1 and COX-2 mRNA were significantly up-regulated after LPS+PGE₂ treatment at 8 and 12 h, and the COX-1 and COX-2 protein were significantly up-regulated (P<0.05). There were no significant effect on the expression of COX-1 and COX-2 mRNA after PGF_2α and LPS present or absent treatment (P>0.05), only significantly increased the expression of COX-1 mRNA after PGF_2α treatment at 8 and 12 h (P<0.05). The expression of COX-1 and COX-2 mRNA were induced after PGE₂and PGF_2α combination treatment group compared with that in PGE₂, PGF_2α and LPS treatment groups.

Key words: bovine endometrial epithelial cells; cyclooxygenase-1(COX-1); cyclooxygenase-2(COX-2); prostaglandin; lipopolysaccharide (LPS)

中图分类号:

S814.8

刘博, 沈媛, 毛伟, 高瑞峰, 曹金山. 前列腺素E₂和F_2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响[J]. 《中国畜牧兽医》, 2017, 44(9): 2716-2723.

LIU Bo, SHEN Yuan, MAO Wei, GAO Rui-feng, CAO Jin-shan. Effect of Prostaglandin E₂ and Prostaglandin F_2α on Cyclooxygenase-1 and Cyclooxygenase-2 Expression in Lipopolysaccharide-treated Bovine Endometrial Epithelial Cells[J]. , 2017, 44(9): 2716-2723.

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前列腺素E₂和F_2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响

Effect of Prostaglandin E₂ and Prostaglandin F_2α on Cyclooxygenase-1 and Cyclooxygenase-2 Expression in Lipopolysaccharide-treated Bovine Endometrial Epithelial Cells

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