东北雨蛙（Hyla japonica）皮肤中新型镇痛肽的发现及其功能研究

发布时间：2018-02-24 07:39

本文关键词： 两栖类皮肤多肽镇痛抗炎　出处：《南京农业大学》2014年硕士论文　论文类型：学位论文

【摘要】：两栖类的皮肤中含有种类繁多、功效复杂的生物活性物质,这些活性物质可用于抵御环境中有害因子的侵犯。两栖类的主要栖息方式有四种：水栖、半陆栖、陆栖和树栖,而大多数雨蛙属两栖类是营树栖生活,且它们一般栖息于水边的植物丛及林木上。在这样的生活环境中它们时刻都可能遭遇更多源于生物及非生物因素的伤害,如：天敌、虫类叮咬、微生物感染及刺伤等等,皮肤作为两栖类生命的第一道防线,在适应生存中固然起着非常重要的作用。雨蛙属绿色或棕色的皮肤不仅使它们更能适应丛林生活,而且皮肤中的一些化学组分对它们的生存也起着至关重要的作用。在本研究中,我们首次从东北雨蛙皮肤中发现了两种新型镇痛多肽,并命名为analgesin-HJ和analgesin-HJ(I5T),它们分别是由5个氨基WPVI-NH2和FWPVT-NH2组成的小肽,且分子量分别为659.84 Da和647.78 Da。analgesin-HJs是由171个氨基酸组成的前体肽序列编码的,其中含有9个重复的analgesin-HJ拷贝和一个analgesin-HJ(I5T)拷贝。然后,我们对analgesin-HJs可能具有的镇痛及抗炎症活性进行了研究,我们通过四种重要的镇痛动物模型对analgesin-HJs的镇痛作用进行研究检测,这四种镇痛动物模型包括小鼠醋酸扭体实验、小鼠福尔马林实验、小鼠光辐射热摆尾实验及热板实验,结果发现analgesin-HJs具有与阳性对照组吗啡相当的镇痛活性。多肽.analgesin-HJ在角叉菜胶诱导的小鼠足趾肿胀模型中的作用同样验证了analgesin-HJ抗炎止痛的效果。除此之外,通过用LPS刺激诱导小鼠巨噬细胞RAW 264.7产生前炎症因子的实验中,我们发现analgesin-HJ具有抑制小鼠巨噬细胞中前炎症因子释放的作用。在对analgesin-HJ镇痛机制的初步研究中,我们发现analgesin-HJ没有明显的阿片受体激动剂活性,其镇痛作用机制还有待进一步深入研究。analgesin-HJs易于合成、运输及储存,加上它们较强的镇痛效果,analgesin-HJs可能为未来开发新型止痛剂提供研究基础。
[Abstract]:The skin of amphibians contains a wide variety of complex bioactive substances that can be used to protect against harmful factors in the environment. There are four main habitats for amphibians: aquatic, semi-terrestrial, terrestrial and arboreal. Most of the amphibians of the genus Rana are tree-dwelling, and they generally live on waterfront vegetation and trees. In such living environments, they are likely to suffer more damage from biological and abiotic factors, such as natural enemies. Insect bites, microbial infections, stab wounds, and so on. Skin, as the first line of defense for amphibian life, plays a very important role in adapting to survival. The green or brown skin of Rana not only makes it more adaptable to jungle life, Some chemical components in the skin also play an important role in their survival. In this study, we found two new analgesic peptides for the first time from the skin of the Northeastern Rana Rana. They are small peptides composed of five amino WPVI-NH2 and FWPVT-NH2, and their molecular weights are 659.84 Da and 647.78 Da.analgesin-HJs respectively, which are encoded by 171 amino acids. There were nine duplicate copies of analgesin-HJ and one copy of analgesin-HJN I5T. Then we studied the possible analgesic and anti-inflammatory activities of analgesin-HJs. We studied the analgesic effects of analgesin-HJs through four important analgesic animal models. The four animal models of analgesia include acetic acid writhing test, formalin test, light radiation tail test and hot plate test. The results showed that analgesin-HJs had the same analgesic activity as that of morphine in positive control group. The effect of polypeptide. Analgesin-HJ on mice toe swelling induced by carrageenin also confirmed the anti-inflammatory and analgesic effect of analgesin-HJ. In the experiment of inducing mouse macrophage RAW 264.7 to produce proinflammatory factor by LPS stimulation, we found that analgesin-HJ can inhibit the release of proinflammatory factor in mouse macrophage. We found that analgesin-HJ has no obvious opioid receptor agonist activity, and its analgesic mechanism needs to be further studied. The analgesin-HJs are easy to synthesize, transport and store. Combined with their strong analgesic effect, analgesin-HJs may provide a basis for the development of new painkillers in the future.
【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965;Q516

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