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首页> 外文期刊>Fish & Shellfish Immunology >Signal transduction of the prophenoloxidase activating system of prawn haemocytes triggered by CpG oligode oxynucleotides
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Signal transduction of the prophenoloxidase activating system of prawn haemocytes triggered by CpG oligode oxynucleotides

机译：CpG寡核苷酸氧核苷酸触发对虾血细胞前酚氧化酶激活系统的信号转导

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Intracellular phenoloxidase (PO) activity in haemocyte lysate supernatant (HLS) of giant freshwater prawn (Macrobrachium rosenbergii) was shown to be enhanced by CpG oligodeoxynucleotide (ODN) 2006, but not by so-ODN13. When haemocytes were treated in vitro with 50 mug/ml of ODN2006 for 30 min, the increases in both intra- and extracellular stimulated PO activity (POs) and extracellular total PO activity (POT) and the reduction Of POT suggest that the PO activity of haemocytes is enhanced by ODN2006 stimulation, but new prophenoloxidase (proPO) is not synthesised. In an attempt to determine which signal transduction pathway is involved in the activation of the proPO system, haemocytes were separately treated with activators or inhibitors of specific signalling components. The results show that there was an increase in both intra- and extracellular POT of haemocytes treated with sodium fluoride (a G-protein activator); the addition of phosphokinase A (PKA)-activating 8-bromo-cAMP to haemocytes only increased intracellular POT, and the addition of either phorbol-12-myristate-13-acetate (PMA; a phosphokinase C (PKC) activator) or caffeine (a phosphodiesterase inhibitor) only increased extracellular POT. When PMA-stimulated haemocytes were treated with chelerythrine (a PKC inhibitor), the induced extracellular POT was significantly reduced. Furthermore, the study of ODN2006-stimulated haemocytes treated with chelerythrine or palmitoyl-DL-carnitine (a PKC inhibitor) showed that the enhancement effects of ODN2006 on the intra- and extracellular POs and extracellular POT were significantly decreased. ODN-stimulated haemocytes treated with genistein (an inhibitor of protein tyrosine kinase) showed a further increase in extracellular POT, but the other PO activities remained the same as those of the ODN-stimulated group. These results suggest that the activation of the proPO system of prawn haemocytes, including degranulation and PO activity, is induced by ODN2006 via a PKC-activating signalling pathway, but negatively regulated via the tyrosine kinase pathway.
机译：CpG寡脱氧核苷酸（ODN）2006可以增强巨型淡水虾（罗氏沼虾）的血细胞裂解物上清液（HLS）中的细胞内酚氧化酶（PO）活性，但so-ODN13则不能。当用50杯/毫升的ODN2006体外处理血细胞30分钟时，细胞内和细胞外刺激PO活性（POs）和细胞外总PO活性（POT）均增加，而POT降低则表明ODN2006刺激增强了血细胞，但未合成新的前酚氧化酶（proPO）。为了确定哪个信号转导途径参与了proPO系统的激活，血细胞分别用特定信号成分的激活剂或抑制剂处理。结果表明，用氟化钠（一种G蛋白激活剂）处理的血细胞的细胞内和细胞外POT均增加;向血细胞中添加激活磷酸激酶A（PKA）的8-bromo-cAMP仅会增加细胞内POT，并且添加phorbol-12-肉豆蔻酸酯13-乙酸酯（PMA;磷酸激酶C（PKC）激活剂）或咖啡因（磷酸二酯酶抑制剂）只会增加细胞外POT。当用白屈菜红碱（一种PKC抑制剂）处理PMA刺激的血细胞时，诱导的细胞外POT显着降低。此外，用白屈菜红碱或棕榈酰-DL-肉碱（一种PKC抑制剂）处理ODN2006刺激的血细胞的研究表明，ODN2006对细胞内和细胞外POs和细胞外POT的增强作用显着降低。用染料木黄酮（蛋白酪氨酸激酶抑制剂）处理的ODN刺激的血细胞显示细胞外POT进一步增加，但其他PO活性与ODN刺激的组相同。这些结果表明，ODN2006通过PKC激活信号途径诱导了对虾血细胞proPO系统的激活，包括脱粒和PO活性，但通过酪氨酸激酶途径对其负调控。

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