摘要
目的初步分离中华仙影海葵溶细胞毒素（CereussinensisVerrillcytolytictoxins，CCT）并探讨其体外溶细胞活性及影响因素。方法采用冻融、匀浆、水浸和离心的方法获取海葵粗毒，通过硫酸铵盐析和Sepha—dexG-100凝胶色谱法将海葵粗毒60％～90％盐析段分为5个组分，检测5个组分溶细胞活性并收集活性较强的第1组分（CCT）；测定其半数溶细胞分数（CU50）、最适孵育温度及最适pH，探讨EDTA和金属离子对溶细胞活性的影响。结果经硫酸铵盐析和分子筛处理后，中华仙影海葵溶细胞活性成分主要集中在第1组分（CCT），CCT溶细胞活性的CUl0为7．76μg·mL-1、最适孵育温度为40℃、最适pH为8；体外实验表明ED-TA和金属离子对CCT溶细胞活性有较大影响。结论通过硫酸铵盐析和凝胶色谱，初步分离得到中华仙影海葵溶细胞活性组分，并测得其主要性质参数，为后续纯化单一溶细胞活性成分打下基础。
Objective To preliminarily isolate cytolytic toxins from Cereus sinensis Verrill and investigate the factors influencing their cytolytic activity. Methods Crude venom from sea anemone was successively obtained by freeze-thaw, homogenate,water extraction and centrifugation. Five parts were isolated from the 60%-90% fraction by ammonium su/fate precipitation and Sephadex G-100 size-exclusion chroma- tograph. The first section （CCT） was shown to be the most potential one by measuring cytolytic activi- ty of them; CU50 ,the best incubation temperature and the best pH of CCT were detected, then the in- fluence of EDTA and metal cations to CCT were investigated. Result CCT was confirmed to be the most potential part by ammonium sulfate precipitation and Sephadex G-100 size-exclusion chromatograph. The CU50 of CCT was 7. 76μg mL-1. the best incubation temperature of CU50 was 40℃ and the best pH of CCT was 8; in vitro cytolytic study of CCT showed that the its cytolytic activity was influenced by EDTA and metal cations mostly. Conclusion The active fraction from Cereus sinensis Verrill was pre- liminarily isolated by ammonium sulfate precipitation and Sephadex G-100 size-exclusion chromato- graph, and the factors influencing cytolytic activity of CCT were detected. This study provides support to discover purified compound subsequently
出处
《中国海洋药物》
CAS
CSCD
北大核心
2013年第5期14-20,共7页
Chinese Journal of Marine Drugs
基金
2012年南通市生物技术和新医药计划项目(AS20120006)资助
关键词
中华仙影海葵
溶细胞活性
活性组分
稳定性
Cereus sinensis Verrill
active component
cytolytic activityl stability
