Rymuszka Anna, Sieroslawska Anna
Department of Physiology and Ecotoxicology, The John Paul II Catholic University of Lublin, Poland.
Neuro Endocrinol Lett. 2011;32 Suppl 1:84-8.
The aim of this study was to determine if cyanoneurotoxin - anatoxin-a (ANTX-a) alters the essential functions of innate immune cells such as free radicals generation in phagocytic cells and phagocytosis.
In the experiments pure ANTX-a was used at concentrations of 0.01, 0.05, 0.1 and 1 µg/ml RPMI-1640 medium. Phagocytes were isolated from carp blood and pronephros. Relative changes in intracellular total free radical presence in fish phagocytes were monitored using a fluorescent probe, dichlorodihydrofluorescin DiOxyQ (DCFH-DiOxyQ) which detects hydrogen peroxide (H2O2), nitric oxide (NO), peroxyl radical and peroxynitrite anion. Phagocytic activity of fish leukocytes was analyzed with a Vybrant phagocytosis assay kit.
The H2O2 level generated in response to ANTX-a at the highest used concentration was significantly suppressed in pronephros but not in blood phagocytes. Moreover, it was observed that generation of superoxide radicals and nitrite formation was significantly increased in blood and pronephros phagocytes after incubation with lower concentrations of the neurotoxin. The phagocytosis of fish leukocytes was significantly reduced at the two highest used toxin concentrations (0.1 and 1 µg/ml medium).
This findings suggests that ANTX-a could change innate immunity and reduced adaptive immunity after stress induced by cyanobacterial blooms.
本研究旨在确定氰神经毒素——anatoxin-a（ANTX-a）是否会改变先天免疫细胞的基本功能，如吞噬细胞中自由基的产生和吞噬作用。
在实验中，使用浓度为0.01、0.05、0.1和1μg/ml的RPMI-1640培养基中的纯ANTX-a。从鲤鱼血液和前肾中分离吞噬细胞。使用荧光探针二氯二氢荧光素二氧喹啉（DCFH-DiOxyQ）监测鱼类吞噬细胞内总自由基存在的相对变化，该探针可检测过氧化氢（H2O2）、一氧化氮（NO）、过氧自由基和过氧亚硝酸盐阴离子。用Vybrant吞噬分析试剂盒分析鱼类白细胞的吞噬活性。
在最高使用浓度下，ANTX-a诱导产生的H2O2水平在前肾中显著受到抑制，但在血液吞噬细胞中未受抑制。此外，观察到在较低浓度神经毒素孵育后，血液和前肾吞噬细胞中超氧自由基的产生和亚硝酸盐的形成显著增加。在最高的两种使用毒素浓度（0.1和1μg/ml培养基）下，鱼类白细胞的吞噬作用显著降低。
这一发现表明，ANTX-a可能会改变先天免疫，并在蓝藻水华引起的应激后降低适应性免疫。
