虾夷马粪海胆体腔液中调理素样分子的研究  

Study of opsonin-like molecule from coelomic fluid of Strongylocentrotus intermedius

作　　者：张颖[1] 李丹彤[1] 常亚青[1] 宋坚[1] 叶仕根[1] 

机构地区：[1]大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《水产学报》2014年第5期684-690,共7页Journal of Fisheries of China

基　　金：辽宁省教育厅团队项目(2008T021)

摘　　要：为研究虾夷马粪海胆体液免疫机理,测定其体腔液吞噬细胞在不同介质中对不同处理的酵母细胞的吞噬作用。对其无细胞体腔液、从调理及非调理酵母细胞分离出的成分进行SDS-PAGE分析,同时对无细胞体腔液进行透析、SephadexG-200凝胶层析和SDS-PAGE分析,在等渗缓冲液和酵母细胞吸附的无细胞体腔液中,测定不同调理素样分子浓度和不同反应时间时吞噬率的变化。结果显示,吞噬细胞在3种介质:等渗缓冲液、酵母细胞吸附的无细胞体腔液和无细胞体腔液中对非调理酵母细胞吞噬率之比为1.00∶1.11∶1.94,在等渗溶液中对调理酵母细胞的吞噬率是非调理酵母细胞的1.61倍;从调理酵母细胞分离出的成分在SDSPAGE中显示一条调理素样分子带,分子量为250.62 ku。经透析和SephadexG-200凝胶层析后,测得纯化的调理素样分子的分子量为250.62 ku;在所测试的浓度范围内,调理素样分子浓度越大,吞噬活性越高,且5.0和10.0μg/mL组在各个测试时间均比对照组高。在等渗缓冲液中,反应30 min时,10.0和5.0μg/mL组与对照组均有极显著差异(P<0.01),在酵母细胞吸附的无细胞体腔液中,反应30 min时,10.0μg/mL组与对照组有显著差异(P<0.05)。结果表明,在虾夷马粪海胆体腔液中发现一种调理素样分子,分子量约为250.62 ku,一定浓度的调理素样分子能显著提高虾夷马粪海胆体腔液吞噬细胞的吞噬活性。In order to study the immune mechanism of coelomic fluid of Strongylocentrotus intermedius,we researched the phagocytosis activity in different media and different treatments of yeast cell. Then we analysed the non-cell coelomic fluid and ingredient which was isolated from regulating yeast cells and nonregulating yeast cells with SDS-PAGE,and the non-cell coelomic fluid was dialysed,SephadexG-200 gel chromatography,and SDS-PAGE; In isotonic buffer and cell-free coelomic fluid by yeast cell adsorption,we determined the phagocytosis effected by opsonin-like molecule of different concentration and different time. The result showed that isotonic buffer,cell-free coelomic fluid by yeast cell adsorption and non-cell coelomic fluid all can regulate phagocytosis,and the ratio was 1. 00 ∶ 1. 11 ∶ 1. 94,and the phagocytosis rate of regulating yeast cells was 1. 61 times of non-regulating yeast cells in isotonic buffer; There was a newopsonin-like molecular band from ingredient which was isolated from regulating yeast cells,and the molecular weight is 250. 62 ku. We also found the molecular weight is 250. 62 ku after dialysis,SephadexG-200 gel chromatography; In tested concentration range,the greater the opsonin-like concentration,the higher the phagocytosis and 5. 0 and 10. 0 μg / mL groups were higher than control group in each test time. In isotonic buffer,10. 0 and 5. 0 μg / mL groups had very significant difference（ P〈 0.01） compared to control group in 30 min after reaction. In cell-free coelomic fluid by yeast cell adsorption,10. 0 μg / mL group had significant difference compared to control group（ P〈0. 05） in 30 min after reaction. These results indicate that there is a kind of newopsonin-like molecule in coelomic fluid of S. intermedius and the molecular weight is 250. 62 ku which can promote phagocytosis.

关 键 词：虾夷马粪海胆 吞噬活性 调理素样分子 吞噬细胞 

分 类 号：Q175[生物学—水生生物学] S917.4[生物学—普通生物学]