Premyslova M, Chisaka H, Okamura K, Challis J R G
CIHR Group in Development and Fetal Health, Department of Physiology and Obstetrics, Gynecology and Medicine, University of Toronto, Toronto, Ontario, Canada M5S1A8.
Placenta. 2006 Jun-Jul;27(6-7):576-86. doi: 10.1016/j.placenta.2005.07.005. Epub 2005 Sep 22.
PGE2 is involved in initiation and progression of labor in many species. Biosynthesis of PGE2 is mediated by cyclooxygenases (COX) and prostaglandin E synthases (PGES). mPGES-1 and COX-2 form an inducible pathway for PGE2 production in many cell systems. In this study we investigated whether mPGES-1 is involved in cytokine induced PGE2 biosynthesis in human trophoblast cells. We have evaluated the cellular and intracellular co-localization of mPGES-1 and COX-2, as well as cPGES and COX-1 in human trophoblast cells by dual immunofluorescent staining. The effect of IL-1beta on mPGES-1 and COX-2 co-localization, such as would occur with infection, and the regulatory effects of pro-inflammatory cytokines IL-1beta and TNF-alpha on transcriptional activity of mPGES-1 and COX-2 in these cells were also studied. We found that in cultured unstimulated trophoblasts, some cells expressed predominantly either mPGES-1 or COX-2, though there were cells co-expressing both enzymes. With IL-1beta treatment, mPGES-1 and COX-2 became more consistently co-localized. mPGES-1 was not transcriptionally co-induced with COX-2 by the cytokine treatment. We conclude that mPGES-1 is not involved in the inducible COX-2 mediated pathway for PGE2 biosynthesis at the transcriptional level, however, the treatment with IL-1beta results in a higher degree of co-ordination of the mPGES-1 and COX-2 protein immunolocalization, eliciting PGE2 synthesis.
前列腺素E2(PGE2)参与了许多物种分娩的启动和进展。PGE2的生物合成由环氧化酶(COX)和前列腺素E合酶(PGES)介导。微粒体前列腺素E合酶-1(mPGES-1)和COX-2在许多细胞系统中形成了一条诱导性的PGE2生成途径。在本研究中,我们调查了mPGES-1是否参与细胞因子诱导的人滋养层细胞中PGE2的生物合成。我们通过双重免疫荧光染色评估了mPGES-1和COX-2以及胞质型前列腺素E合酶(cPGES)和COX-1在人滋养层细胞中的细胞内和细胞间共定位情况。还研究了白细胞介素-1β(IL-1β)对mPGES-1和COX-2共定位的影响(如感染时会出现的情况),以及促炎细胞因子IL-1β和肿瘤坏死因子-α(TNF-α)对这些细胞中mPGES-1和COX-2转录活性的调节作用。我们发现,在未受刺激的培养滋养层细胞中,一些细胞主要表达mPGES-1或COX-2,不过也有细胞同时表达这两种酶。用IL-1β处理后,mPGES-1和COX-2的共定位变得更加一致。细胞因子处理并未使mPGES-1与COX-2在转录水平上共同诱导。我们得出结论,在转录水平上,mPGES-1不参与诱导性COX-2介导的PGE2生物合成途径,然而,用IL-1β处理会导致mPGES-1和COX-2蛋白免疫定位的更高程度协调,从而引发PGE2合成。