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Dolmatova L. S. Eliseykina M. G. Timchenko N. F. Kovaleva A. L. Shitkova O. A. 《中国海洋湖沼学报》2003,21(4):293-304
Pure fraction (92%–95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38%)-FPMC- of the holothurianEupentacta fraudatrix' (Holothuroidea, Dendrochirota) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified
by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin ofYersinia pseudotuberculosis (TST) at different concentrations (0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation.
In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction
after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively.
Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared
to that under toxin treatment alone. TST stimulated superoxide dismutase activity in concentration-dependent manner (maximum
at 0.5 μg/ml concentration in FP) after 24 h treatment, and this stimulation was prevented by a commercial catalase. Plant
lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin
stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously,
ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in
holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production
by these cells may be one of the mechanisms of antibacterial protection of holothurians.
This work was financially supported by the Russian Foundation for Fundamental Research Grant (No. 00-04-48949). 相似文献
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Dolmatova L.S. Eliseykina M.G. Timchenko N.F. Kovaleva A.L. Shitkova O.A. 《中国海洋湖沼学报》2003,21(4):293-304
Pure fraction (92% - 95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38 % )-FPMC- of the holothurian Eupentacta fraudatrix ( Holothuroidea,Dendrochirota ) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin of Yersinia pseudotuberculosis (TST) at differentconcentrations ( 0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in ccncentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 htreatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians. 相似文献
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采用CCM、HSM和L-15N 3种培养液对刺参(Apostichopus japonicas)的体腔细胞进行了体外原代培养,并用MTT还原法对细胞的体外存活力进行测定,结果显示:CCM和HSM培养的体腔细胞,在体外分别存活至3 d和6 d大量死亡;L-15N培养液可使细胞在1周内保持存活.用刺参病原菌灿烂弧菌的胞外产物与刺参体外培养细胞共培育,发现对体腔细胞有毒性作用,半致死蛋白质浓度(IC_(50))为27.6 μg/mL. 相似文献
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