虾夷马粪海胆(Strongylocentrotus intermedius)体腔液的酚氧化酶活性分析

Analysis of Phenoloxidase Activity in the Coelomic Fluid of Strongylocentrotus intermedius

分别测定虾夷马粪海胆血浆和体腔细胞溶解上清液(CLS)中的酚氧化酶(PO)活性以及不同刺激物对其酚氧化酶原系统(proPO)的激活效果,并初步分析了患病虾夷马粪海胆体腔液中酚氧化酶活性的变化。结果显示:虾夷马粪海胆体腔液的PO主要分布于血浆中,且个体差异较大,血浆中PO活性为48.28±6.69U,CLS中PO活性为2.24±1.81 U;proPO存在于体腔细胞中,含量较少,其被1 mg/mL Trypsin及100 mmol/LMgCl2激活后,PO活性分别提高2.24 U和7.86 U,LPS、SDS、CaCl2和甲醇等对酚氧化酶原无明显激活效果;虾夷马粪海胆感染细菌后体腔液的酚氧化酶活性为12.90±2.03 U,相较于健康海胆(30.63±2.21 U)显著降低(P<0.05),而在100 mmol/L MgCl2作用下酚氧化酶活性提高4.19±1.96 U,相较于健康海胆(2.76±1.15U)明显升高(P<0.05)。研究结果为虾夷马粪海胆及其他棘皮动物的免疫防御机制的深入研究提供了基础。

Phenoloxidase（PO） activity and proPO induction in plasma and coelomocyte lysate supernatan（CLS） of Strongylocentrotus intermedius was detected,and the effect of disease on the enzyme actives of sea urchin was analyzed.The results showed that PO of the coelomic fluid of S.intermedius was mainly distributed in plasma,and the PO activity in plasma was 48.28±6.69 U,that in CLS was 2.24±1.81 U;and prpPO existed in CLS was at very low level.After being activated by 1 mg/mL Trypsin and 100 mmol/L MgCl2,the PO activity increased 2.24 U and 7.86 U,respectively.The LPS,SDS,CaCl2 and methanol had no obviously inductive effect;and the PO activity of bacteria-infected S.intermedius was 12.90±2.03 U,which declined significantly（P0.05）comparing with that of the healthy ones（30.63 ±2.21 U）.While the PO activity of diseased S.intermedius increased 4.19±1.96 U after induced by 100 mmol/L MgCl2,which increased significantly（P0.05）comparing with that of the healthy ones（2.76±1.15 U）.The above results provide basis for further studies on immune defense mechanism of S.intermedius and other Echinodermata aminals.