【作者】 张文涛；

【导师】 汤鲁宏；

【作者基本信息】 江南大学 ， 微生物与生化药学， 2008， 硕士

【摘要】 霞水母是严重威胁我国渤海和黄海北部的渔业生态资源的一种海洋生态灾害物种,霞水母爆发与赤潮并列为我国海洋生态的两大灾难。为了解决霞水母爆发对我国海洋生态环境的灾难性影响,急需进行霞水母的资源化利用方面的研究,以便增加其附加值,提高渔民的捕捞积极性,确保霞水母的捕捞死亡率大于其自然增长率,逐渐消除其对海洋生态环境的影响。然而,迄今为止,有关霞水母的资源化利用方面的研究尚属空白,国内外迄今未见任何相关报道。胶原蛋白作为细胞外基质中起支持作用的主要成分,具有“Gly-X-Y”的特异性氨基酸组成和螺旋结构,因其还具有抗原性低、可生物降解性强、生物相容性好等优点,故在生物学、医学、食品科学和美容业等领域中具有巨大的研究开发潜力和广阔的应用前景。国内外对胶原蛋白的分离制备主要以哺乳动物皮肤、软骨以及鱼类皮肤等组织为原料,对其研究主要集中在分离制备和生物材料领域的开发应用方面。到目前为止,对腔肠动物胶原蛋白的研究尚较少,霞水母来源胶原蛋白的分离纯化及其生理活性方面的研究迄今国内外未见任何报道。为了实现霞水母的资源化开发利用,本文在国内外首次就霞水母中含有的胶原蛋白进行了研究,建立了霞水母胶原蛋白的分离制备和纯化方法,并就其是否对类风湿性关节炎的预防和治疗具有生物活性进行了探索性研究,为将霞水母中所含胶原蛋白作为天然海洋胶原蛋白资源加以开发利用,运用环境经济理念,消除这一海洋生态灾难,提供了工作基础和实验依据。本文用酶法从霞水母中提取得到了胶原蛋白。根据聚丙烯酰胺凝胶电泳、变性聚丙烯酰胺凝胶电泳图谱推测霞水母胶原蛋白可能是由三条分子大小在66.2 KDa-116 KDa之间不同亚基组成的三聚体。样品中蛋白质含量为98.0%。本实验所测得胶原蛋白的羟脯氨酸含量为10.54%,而糖无检出,表明胶原蛋白纯度较高。急性毒性试验表明,在剂量为15000 mg/kg霞水母胶原蛋白的情况下,未出现小鼠死亡,也未有明显毒性反应。本实验对霞水母胶原蛋白口服治疗大鼠佐剂型关节炎进行了初步探索。发现口服霞水母胶原蛋白可有效减轻AA大鼠的关节肿胀度,显著降低AA大鼠血清MDA、NO水平,提高血清SOD的活性。通过这些指标,可以推测胶原蛋白对大鼠佐剂型关节炎可能有一定的治疗效果。更多还原

【Abstract】 Cyanea nozakii is a kind of harmful jellyfish for marine ecosystem which bloom out in the Bohai Sea and Northern Yellow Sea of China. In fact, the jellyfish bloom and Harmful Algal Bloom are top two ecological disaster of Chinese marine ecosystem, threatening the fishery industry. In order to eliminate threaten of jellyfish bloom on the marine fishery industry of our country. It is desiderated to undertake the basic research about how to use it as a kind of new marine resource. From the environmental economic point of view, it is the best solution try to find the way to utilize it. About how to use it as a kind of resource, however, we know nothing. Nothing has been reported about the basic knowledge and information about basic components, as well as their function up to now.Collagen is a major component of extracellular matrix (ECM), and has a unique three-dimensional structure with a unique amino acid composition. Owing to wonderful properties of its low-antigenicity, easy-biodegradability and excellent biocompatibility, collagen is used more and more widely in areas of biological material development, food industry, cosmetic manufacture, and medical production as well.Recently, there have been many interests in establishing efficient techniques to isolate collagen from skin of fish, and skin, cartilage and soon from mammals, and to apply them in development of various kinds of biological materials. However, there has been no report about the collagen from the Cyanea nozakii, as well as its biological functions on adjuvant arthritis treatment.For the first time, the collagen from Cyanea nozakii has been investigated comprehensively. The isolation and purification methods have been established, and the potential physiologic activity of the collagen from Cyanea nozakii on the prevention and treating of adjuvant arthritis has also been explored. The data and results collected and recorded on this thesis can be used as the basic information for the further research and development of Cyanea nozakii, as a kind of new marine conllagen resources.The collagen from Cyanea nozakii has been extracted by enzymatic method. The collagen sample obtained was examined by PAGE and SDS-PAGE, which indicate that collagen from Cyanea nozakii may have the chain composition of a heterotrimer and the subunits weighed from 66.2 KDa to 116 KDa. Respectively, the percentage of hydroxyl-proline in the collagen after purification was 10.54%, the concentration of the protein was 98.0%, and the concentration of the polysaccharide was no detection.The acute toxicity experiment showed that injecting big dose, mice didn’t die and hadn’t obvious toxicity reaction. In a word, the collagen from Cyanea nozakii has no toxicity, which security dose was 15000 mg/kg.In this study, therapeutic effect of Cyanea nozakii collagen by oral administration on adjuvant arthritis in rats was initiatory explored. Cyanea nozakii collagen at different concentration could ameliorate the adjuvant induced arthritis, suppress the concentrations of NO and MDA in the blood serum and increase the activity of SOD in the blood serum. From these indexes can presume that, Cyanea nozakii collagen is probably ameliorating the adjuvant induced arthritis.更多还原