华支睾吸虫来源极性差异化合物的高效薄层色谱免疫分析  

Using high-performance thin-layer chromatographic and immunological techniques to identify compounds with varying polarities fromClonorchis sinensis

作　　者：杨庆利[1,2] 申继清[3] 蒋智华[2] 陈颖丹[1] 周晓农[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾,血吸虫病和丝虫病合作中心,上海200025 [2]广西壮族自治区疾病预防控制中心,广西南宁530028 [3]广西医科大学寄生虫学教研室,广西南宁530021

出　　处：《中国病原生物学杂志》2014年第9期816-819,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.31260221);广西壮族自治区自然科学基金项目(No.2010GXNSFB013066)

摘　　要：目的探索华支睾吸虫ESPs、虫卵和虫体水溶性组分中生物大分子极性和抗原性的差异并寻找可行的分离方法。方法用HPTLC法分离华支睾吸虫ESPs、虫卵和虫体的水溶性组分,同时结合免疫染色法研究极性差异组分的抗原性。结果用有机溶剂组合可将ESPs、虫卵和虫体水溶性组分中极性强弱差异的化合物分离开。氯仿︰甲醇︰水(11︰9︰2)溶剂组合对弱极性化合物各组分有较好的分离效果。与虫卵相比较,ESPs和虫体的弱极性组分较复杂。不能被有机溶剂展开的强极性组分的抗原性较强,未检测出弱极性组分的抗原性反应。通过有机溶剂提取的弱极性组分可完全在水中复溶并被有机溶剂重新展开。结论华支睾吸虫来源的水溶性组分中的弱极性化合物成分能通过有机溶剂组合进行有效分离。这为分离华支睾吸虫来源的脂类等弱极性生物大分子以及研究其病原相关分子模式的性质奠定了基础。Objectives To examine the polarity and antigenicity of biological macromolecules in water-soluble components from excretory-secretory products（ESPs）,eggs,and adult worms of Clonorchis sinensis and to explore suitable methods of separating those components. Methods Water-soluble components were separated from ESPs,eggs,and adult worms using high-performance thin-layer chromatography（HPTLC）.Differences in the antigenicity of compounds with varying polarities were analyzed using immunostaining with mouse antiserum against C.sinensis. Results Compounds with varying polarities were separated from the water-soluble components of ESPs,eggs,and adult worms via a combination of organic solvents.A chloroform：methanol：water solvent（11︰9︰2）was rather effective at separating compounds with a weak polarity.ESPs and adult worms had a more complex composition of compounds with a weak polarity than did eggs.Components with a strong polarity could not be extracted with organic solvents but they did appear to be highly antigenic.However,components with a weak polarity did not produce an antigenic reaction.In addition,watersoluble compounds with a weak polarity were extracted with a combination of organic solvents and they were redissolved in water and extracted again using organic solvents. Conclusion Compounds with a weak polarity were effectively separated from the water-soluble components of C.sinensis using a combination of organic solvents.This finding will facilitate the separation of compounds with a weak polarity,such as lipids,and provide a basis for examination of the pathogen-associated molecular patterns（PAMPs）of C.sinensis.

关 键 词：华支睾吸虫 极性 抗原性 HPTLC 病原相关分子模式 

分 类 号：R383.22[医药卫生—医学寄生虫学]