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首页> 外文期刊>Cell and Tissue Research >The prophenoloxidase system is activated during the tunic inflammatory reaction of Ciona intestinalis
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The prophenoloxidase system is activated during the tunic inflammatory reaction of Ciona intestinalis

机译：前肠氧化鞘膜炎性反应期间，前酚氧化酶系统被激活

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摘要

Phenoloxidase (PO) activity was examined in the tunic tissue of Ciona intestinalis following lipopolysaccharide (LPS) intratunic injection. Tunic homogenate supernatant (THS), assayed with the Dopa-MBTH reaction, displayed Ca2+-independent PO activity that was raised by LPS and further enhanced by proteases. Specific inhibitors (tropolone, phenylthiourea, diethylthiocarbamate) supported the specificity of the reaction. Assay with soybean trypsin inhibitor showed that, in the tunic, PO activation with trypsin was not significantly inhibited suggesting that proteases diverse from serine proteases were involved. In vivo experiments were carried out by injecting isosmotic medium or LPS, and THS was assayed for its PO activity. Analysis of variance of the time-course profiles showed that LPS was more effective in activating proPO. To disclose the PO response at the injured site, an assay with Dopa-MBTH was performed in vitro. Quinones were mainly contained in the tunic matrix enriched with inflammatory cells around the injection site. Microscopic observations and immunohistochemistry with anti-CinPO-2 antibodies showed granulocytes and unilocular refractile granulocytes containing PO, whereas few morula cells were stained. In THS zymograms (SDS-polyacrylamide gel electrophoresis), PO activity linked to 90-kDa and 120-kDa bands was observed as an effect of LPS injection, whereas the density of 170-kDa PO was weak. A third presumptive PO enzyme (CinPO-3) containing the CinPO-2 peptide was identified in the recent Ciona genome version. Presumably, LPS stimulated the production and dimerization (120 kDa) of CinPO-3 (66 kDa). Thus, the activated proPO system includes several POs that are distinguishable by size and that are contained and presumably released by tunic inflammatory cells and hemocytes of the pharynx bars.
机译：脂多糖（LPS）鞘内注射后，在Ciona肠的中风组织中检查了酚氧化酶（PO）的活性。用Dopa-MBTH反应测定的外衣匀浆上清液（THS）表现出Ca2 +不依赖的PO活性，该活性由LPS升高并由蛋白酶进一步增强。特定的抑制剂（托酚酮，苯硫脲，二乙基硫代氨基甲酸酯）支持反应的特异性。用大豆胰蛋白酶抑制剂的分析表明，在外衣中，用胰蛋白酶的PO活化没有受到显着抑制，这表明与丝氨酸蛋白酶不同的蛋白酶也参与其中。通过注入等渗培养基或LPS进行体内实验，并测定THS的PO活性。时程分布的方差分析表明，LPS在激活proPO方面更有效。为了公开在受伤部位的PO反应，在体外用Dopa-MBTH进行了测定。醌主要包含在注射部位周围富含炎症细胞的中药基质中。显微镜观察和抗CinPO-2抗体的免疫组织化学分析显示，粒细胞和单眼屈光性粒细胞含有PO，而桑细胞很少染色。在THS酶谱图（SDS-聚丙烯酰胺凝胶电泳）中，观察到与LPS注射作用有关的与90-kDa和120-kDa谱带相关的PO活性，而170-kDa PO的密度较弱。在最新的Ciona基因组版本中，发现了第三个推定的PO酶（CinPO-3），该酶含有CinPO-2肽。据推测，LPS刺激了CinPO-3（66 kDa）的产生和二聚化（120 kDa）。因此，活化的proPO系统包括几个PO，这些PO在大小上是可区分的，并且被咽部条的炎性细胞和血细胞所包含并且可能被释放。

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  • 来源
    《Cell and Tissue Research》 |2008年第3期|481-492|共12页
  • 作者

    M. Cammarata; V. Arizza; C. Cianciolo; D. Parrinello; M. Vazzana; A. Vizzini; G. Salerno; N. Parrinello;

  • 作者单位

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

    Marine Immunobiology Laboratory Department of Animal Biology University of Palermo Via Archirafi 18 Palermo Italy;

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  • 正文语种 eng
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  • 关键词

    Phenoloxidase; Hemocyte; Tunic; Inflammation; Lipopolysaccharide; SDS-polyacrylamide gel electrophoresis; Ciona intestinalis (Tunicata);

    机译：苯酚氧化酶;血细胞;中药;炎症;脂多糖;SDS-聚丙烯酰胺凝胶电泳;小肠虫（Tioncata）;

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