蛇毒中两种活性成份——中华眼镜蛇（Naja naja atra)蛇毒神经生长因子及烙铁头（Trimeresurus mucrosquamatus）蛇毒磷脂酶A_2样肌肉毒的研究

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	蛇毒中两种活性成份——中华眼镜蛇（Naja naja atra)蛇毒神经生长因子及烙铁头（Trimeresurus mucrosquamatus）蛇毒磷脂酶A_2样肌肉毒的研究
	徐天瑞
学位类型	博士
导师	熊郁良
	1998-07
学位授予单位	中国科学院研究生院
学位授予地点	北京
关键词	神经生长因子中华眼镜蛇蛇毒棉酚生殖磷酯酶a_2 烙铁头蛇毒肌肉毒血小板聚集抑制剂肝素
摘要	本文采用与前人不同的新方法，利用离子交换、分子筛和快速蛋白质液相色谱（FPLC）从中华眼镜蛇华南亚种（Naja naja atra）蛇毒中分离纯化一种神经生长因子（NGF），其SDS-PAGE分子量为13500，等电点为7.2。与小鼠颌下腺神经生长因子相比中华眼镜蛇毒神经生长因子具有较低的活性。其N端蛋白质序列与其它眼镜蛇属蛇毒神经生长因子一致。用PCR方法扩增了该蛋白的编码基因并进行了序列测定。大鼠腹腔注射中华眼镜蛇毒神经生长因子可增加其附睾尾部精子的活力，提高交配雌鼠的怀孕率和胎仔数，并可以拮抗棉酚对睾丸的破坏作用，降低因棉酚损伤而升高的血清LH、FSH水平。这些结果提示神经生长因子能改善雄性大鼠的生殖功能，具有明显的促生育作用。在从烙铁头（Trimeresurus mucrosquamatus）蛇毒中分离神经生长因子的过程中，我们发现了一个新的碱性磷酯酶A_2样肌肉毒，命名为TMPB，其SDS-PAGE分子量为16000，等电点为9.2。其N端24个氨基酸序列与烙铁头属的其它磷酯酶A_2同源性较低。TMPB没有明显的水解卵磷脂活性，但它的肌肉毒和血小板聚集抑制活性却极强，其肌肉毒和血小板聚集抑制活性可被肝素所抑制。
其他摘要	A nerve growth factor (NGF) was isolated from the venom of Naja naja atra by ion exchange chromatography, gel filtration and Fast Protein Liquid Chromatography (FPLC). The NGF monomer molecular weight was estimated to be 13,500 by SDS-PAGE, with an isoelectric point being 7.2. The purified NGF showed lower activity than mouse NGF. The mature NGF coding gene was amplified by polymerase chain reaction and sequenced. NGF could improve the sperm motility of cauda epididymal of male rats, increase the pregnant rate and fetus number of mated female rats. The serum hormone level of LH and FSH of male rats administrated with NGF was lower than that of male rats administrated with NGF + gossypol, this together with the histological sections of tests and epididymal suggested that NGF could improve the reproductive function of male rats and reduce the destructive effects of gossypol on testes. When we isolating nerve growth factor from the venom of Trimeresurus mucrosquamatus, we purified A new basic phospholipase A_2-like myotoxin, designated TMPB, from the venom of T. mucrosquamatus. The N-terminal sequence of 24 amino acid residues was determined, the similarity was rather low comparing with the sequence of PLA_2s previously purified from the same genus. The molecular weight was estimated to be 16,000 by SDS-PAGE, the isoelectric point was determined to be 9.2 by isoelectric focusing electrophoresis. The TMPB showed no detectable hydrolytic activity, while exhibited strong myotoxicity and platelet aggregation inhibiting activity. The myotoxicity and platelet aggregation inhibiting activity could be obviously inhibited by heparin.
语种	中文
文献类型	学位论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/6201
专题	其他
推荐引用方式 GB/T 7714	徐天瑞. 蛇毒中两种活性成份——中华眼镜蛇（Naja naja atra)蛇毒神经生长因子及烙铁头（Trimeresurus mucrosquamatus）蛇毒磷脂酶A_2样肌肉毒的研究[D]. 北京. 中国科学院研究生院,1998.

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