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Effects Of Tegdma And Hema On The Expression Of Cox-2 And Inos In Cultured Murine Macrophage Cells

机译:Tegdma和Hema对体外培养的小鼠巨噬细胞Cox-2和Inos表达的影响

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Objective. This study is aimed to investigate the effects of triethyleneglycol dimethacry-late (TEGDMA) and 2-hydroxyethyl methacrylate (HEMA) on expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in cultured murine macrophage cell line RAW 264.7. Methods. For mRNA gene expression analysis of COX-2 and iNOS, RAW 264.7 cells were exposed to TEGDMA and HEMA, and mRNA of each gene was observed by using a reverse transcriptase polymerase chain reaction (RT-PCR) assay. The Western blotting method was applied for detection of COX-2 and iNOS proteins extracted from RAW 264.7 cells treated with the resin monomers. Prostaglandin E2 (PGE2) was quantified by immunoassay with commercially available ELISA kits. Results. TEGDMA and HEMA induced the expression of COX-2 mRNA dose-dependently in RAW 264.7 cells. The expression of COX-2 mRNA was up-regulated at 5 h of exposure to both TEGDMA and HEMA, and diminished thereafter. The resin monomers did not affect the expression of iNOS mRNA. Up-regulation of COX-2 protein was confirmed in the cells treated with TEGDMA and HEMA. Production of PGE_2 was also enhanced by TEGDMA. However, HEMA did not affect PGE_2 biosynthesis, although HEMA up-regulated the expression of COX-2 mRNA and protein. Significance. These findings suggest that TEGDMA and HEMA can be a critical factor of inflammation related to resin-based dental biomaterials, and that COX-2 is involved in the inflammatory reaction of the resin monomers.
机译:目的。这项研究旨在调查三乙二醇二甲基丙烯酸酯(TEGDMA)和甲基丙烯酸2-羟乙酯(HEMA)对培养的鼠巨噬细胞细胞系RAW中环氧合酶2(COX-2)和诱导型一氧化氮合酶(iNOS)表达的影响264.7。方法。对于COX-2和iNOS的mRNA基因表达分析,将RAW 264.7细胞暴露于TEGDMA和HEMA,并通过逆转录聚合酶链反应(RT-PCR)分析观察每个基因的mRNA。蛋白质印迹法用于检测从用树脂单体处理的RAW 264.7细胞中提取的COX-2和iNOS蛋白。前列腺素E2(PGE2)通过市售ELISA试剂盒的免疫测定进行定量。结果。 TEGDMA和HEMA诱导RAW 264.7细胞中COX-2 mRNA的表达呈剂量依赖性。在暴露于TEGDMA和HEMA的5小时内,COX-2 mRNA的表达上调,此后降低。树脂单体不影响iNOS mRNA的表达。在用TEGDMA和HEMA处理的细胞中证实了COX-2蛋白的上调。 TEGDMA也提高了PGE_2的产量。尽管HEMA上调了COX-2 mRNA和蛋白的表达,但HEMA并未影响PGE_2的生物合成。意义。这些发现表明,TEGDMA和HEMA可能是与基于树脂的牙科生物材料相关的炎症的关键因素,并且COX-2参与了树脂单体的炎症反应。

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