沙蜇刺丝囊毒素的分离提取及其溶血活性稳定性研究

Study on the extraction and the stability of the hemolytic activity of nematocyst venom from the jellyfish Stomolophus meleagris

目的:研究沙蜇刺丝囊毒素的提取及其溶血活性稳定性的影响因素。方法 :通过自溶法制备沙蜇刺丝囊细胞,利用超声波破碎细胞提取毒素,再将毒素置于不同温度、不同pH值及不同浓度的乙二胺四乙酸(EDTA)、还原型谷胱甘肽(GSH)及二价金属离子溶液中孵育后,加入到人血红细胞溶血反应体系中,测定上述理化因素对沙蜇刺丝囊毒素溶血活性稳定性的影响。结果:成功提取到沙蜇刺丝囊毒素,其半溶血率(HU_(50))约为26.38 mg/L;毒素的溶血活性受温度影响较大,37℃时溶血活性最强;溶血活性在弱酸性条件下较稳定,pH4.8时毒素的溶血活性最大;EDTA和GSH对毒素溶血活性具有稳定作用,而二价金属离子Ca^(2+)、Mn^(2+)、Cu^(2+)、Mg^(2+)、Ba^(2+)和Zn^(2+)能不同程度的抑制其活性。结论:沙蜇刺丝囊毒素具有溶血活性,其活性的稳定性受温度、pH值、EDTA、GSH及二价金属离子影响。

Objective： To study the extraction and factors affecting the stability of the hemolytic activity of nematocyst venom from the jellyfish Stomolophus meleagris. Methods： The nematocysts were prepared using the method of autolysis in sterile sea water, followed by uhrasonic homogenization and extraction of venom. The nematocyst venom was added to the hemolytic sys- tem of human erythrocytes to evaluate the effects of the physical and chemical factors on the stability of the hemolytic activity of the jellyfish Stomolophus meleagris venom after incubation at different temperature, pH and concentrations of ethylenedi- aminetetraacetie acid（EDTA）, reduced glutathione（GSH） and divalent metal ions. Results： The nematocyst venom was extracted successfully from the jellyfish Stomolophus meleagris. Approximately 26.38 mg/L of the venom resulted in 50% hemolysis of the human erythrocytes. Temperature greatly affected hemolytic activity of the venom, with highest hemolytic activity at 37 ℃. Hemolytic ac- tivity of the venom was relatively stable at weak acid, with maximal activity at pH 4.8. Both GSH and EDTA stabilized the hemolytic activity of Stomolophus meleagris venom, while Ca2＋,Mn2＋,Cu2＋,Mg2＋,Ba2＋ and Zn2＋ could inhibit hemolytic activity at certain extent. Conclusions： The venom of Stomolophus meleagris exhibits hemolytic activity, which stability is influenced by some physical and chemical factors, including temperature, pH, EDTA, GSH and divalent metal ions.