中国对虾血细胞体外吞噬活动中活性氧产生的研究

IOCAS-IR > 海洋环流与波动重点实验室

	中国对虾血细胞体外吞噬活动中活性氧产生的研究
其他题名	Studies on ROS production in phagocytosis of Penaeus chinesis hemocyes in vitro
	王宝杰
学位类型	博士
	2002
学位授予单位	中国科学院海洋研究所
学位授予地点	中国科学院海洋研究所
学位专业	海洋生物学
关键词	中国对虾血细胞存活率化学发光法 Nbt还原法
摘要	该文首先对中国对虾血细胞体外短时培养条件进行了摸索,以期为后来在体外研究中国对虾血细胞吞噬活动中ROS的产生建立基础.在上在实验的基础上,该文利用化学发光法对中国对虾血细胞体外吞噬过程中ROS的产生进行了研究,试图了解血细胞吞噬活动中的化学发光现象.最后利用NBT还原法研究了中国对虾血细胞体外吞噬过程中O<'-><,2>的产生化及一些环境污染物如重金属离子和农药对其吞噬活动中O<'-><,2>的产生影响.
其他摘要	Penaeus chinesis is a very important commercial animal on Chinese mari-culture, however, the shrimp industry has always been affected by infectious diseases, mainly of bacterial and viral etiology. The research on the immunity mechanisms of shrimp become an urgent and basic need. We studied the ROS production in phagocytosis of P. chinensis hemocytes in vitro, in order to illuminate the cellular immune mechanisms of shrimp. In this study, We firstly study a culture condition for improving short-term in vitro maintenance of hemocytes from P. chinensis, intending to lay the foundation of research on the ROS production in phagocytosis of P. chinensis hemocytes subsequently. According to cell viability assayed with a trypan blue exclusion test, we established the optimal conditions of culture time, culture medium and culture temperature respectively. The result indicate: a significantly higher percentage of hemocytes viability was obtained following cultured in 30min, CM and SCM solution , 4 ℃ and 27 ℃ conditions compared to other conditions, which show these conditions fit for the in vitro manipulation of chinensis hemocytes. On basis of the above experiments, we study the ROS production in phagocytosis of P. chinensis hemocytes in vitro using a CL method, intending to comprehend the CL phenomenon in phagocytosis. The result indicate: Zymosan A and LPS can both stimulate P. chinensis hemocytes to generate CL response, which show the hemocytes of P. chinensis undergo a respirtory burst in phagocytosis. But the cpm of CL is faint. Finally we studied the O_2~- production and the affect of some environmental contamination on O_2~- production in phagocytosis of P. chinensis hemocytes in vitro using NBT reduction. The result indicate: the hemocytes of/! chinensis will undergo an obvious burst of respiratory and generate O_2~- , when they were stimulated in vitro to begin their phagocytic activity. With the same stimulator (Zymosan A and LPS) under different concentrations, the hemocytes showed different amount production of O~2; Different hemocytes concentrations that were activated with the optimal concention of Zymosan A show different amount production of O_2~-; SODN NEM and Iodoacetamide could produce a inhibition of O_2~- production; 5 kinds heavy metals and 2 kinds pesticides could also inhibite O_2~- production; Individual animals showed a relatively high degree of variability in the production of O_2~-, The amort shrimps showed a relatively low capability of O_2~- production. It comes to the conclusion that the hemocytes of P. chinensis can generate O_2~- etc. ROS to kill their pathogens in the phagocytic defensive reaction. Compared with CL, the NBT reduction is more propitious to be used as the index to determine the prawn immune state.
页数	63
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/1457
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	王宝杰. 中国对虾血细胞体外吞噬活动中活性氧产生的研究[D]. 中国科学院海洋研究所. 中国科学院海洋研究所,2002.

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