【作者】 张宸阁；

【导师】 卢卫红；

【作者基本信息】 哈尔滨工业大学 ， 食品科学， 2011， 硕士

【摘要】 孔石莼是一种广泛分布于西太平洋沿海地区的大型绿藻,资源十分丰富。其具有很高的经济和药用价值,富含蛋白质、氨基酸、糖类等多种营养成分,味道鲜美.孔石莼多糖可增强机体免疫力,具有抗擬血、抗肿瘤、降胆固醇、抗氧化等药用保障功能。实验中对孔石莼多糖进行水提醇沉,并通过去蛋白、高速离心等对粗多糖进行纯化,预处理后通过100ku和30ku超滤膜对孔石莼多糖进行分级纯化,最终将其分为了≧100ku、30~100ku及≦30ku三个不同分子量段的多糖溶液。经小鼠骨髓细胞微核、肝脾脏系数测定、SOD活性测定及MDA含量测定等多项抗辐射实验,对其抗辐射作用效果进行了比较和筛选,最终发现≦30ku分子量段多糖溶液相较于另两种,其对因辐射引起的骨髓微核率的提升有明显抑制作用(P<0.01),对脾脏系数有较明显的升高(P<0.05),且在小鼠辐照后明显提升了其肝脏中SOD的活性并有效降低了其MDA的含量,通过以上实验结果显示,≦30ku分子量段多糖抗辐射作用强。取≦30ku分子量段多糖与铁(Ⅲ)离子进行螯合,得到孔石莼多糖铁(Ⅲ)配合物。能谱显示其铁元素的含量较高,原子分光光度法测得其铁含量为27.4%,通过红外光谱显示,其在850cm-1和650cm-1处有特征峰,显示其铁核为β-FeOOH。对多糖及多糖铁(Ⅲ)进行抗辐射实验后发现,两者均有较强的抗辐射作用,但效果差别不大。对多糖和多糖铁(Ⅲ)辐射实验中的小鼠脾脏进行2-DE蛋白组学研究。分析各组差异点后发现,相对于空白组,辐照对照组中2倍下调的蛋白点有13个,2倍上调的蛋白点有2个;相对于辐照对照组,多糖组中2倍下调蛋白点有5个,2倍上调的蛋白点有14个,多糖铁组中有8个2倍下调的蛋白点,12个2倍上调的蛋白点。从中选取五个差异点进行质谱分析(3502,4705, 5402,5803及6801),发现此五种蛋白分别为78kDa葡萄糖调控蛋白,脯氨酰羟化酶beta,亚基钙网蛋白前体,微管蛋白β5及波形蛋白等。经分析发现,这五种蛋白均与机体免疫及损伤相关,通过对多糖及多糖铁与辐照对照组相比较,发现这两者都在大多蛋白表达上有效果,且其抗辐射效果较好两者差异较小。更多还原

【Abstract】 coast, and its resources are very rich. Ulva pertusa has a high economic and medicinal value, contains protein, amino acids, sugars and other nutrients,who is very delicious.The polysaccharide of Ulva pertusa can enhance immunity, with the proposed anti-blood, anti-tumor, reduce cholesterol, anti-oxidant and other medical support functions.The polysaccharide of Ulva pertusa was extracted by water,and precipitated with ethanol, then separated by centrifuagation, the crude polysaccharide extract was obtained. After pretreatment, the polysaccharide of Ulva pertusa was futher purified by 100ku and 30ku membrane, and then the polysaccharide of Ulva pertusa was divided into different molecular weight polysaccharide solution(≧100ku、30~100ku and≦ 30ku). The radioprotective ability of different molecular weight polysaccharide solution were studied using micronucleus assay of mice bone marrow cells, and determination of the spleen index, the activity of SOD and the level of malondialdehyde (MDA). Eventually found the polysaccharide solution less than 30ku showed best ability of radioprotection, it reduced the radiation induced frequency of micronuclei(P<0.01), and increasing the spleen index (P<0.05) and the activity of SOD in the liver after irradiation in mice significantly. The radiation-induced decrease of MDA in the liver by pretreatment wtth polysaccharide (≦30ku). The results show that polysaccharide(≦30ku) possess effitive antiradiation activity.The polysaccharide of Ulva pertusa(≦30ku) and Fe(Ⅲ) chelation,and got Ulva pertusa polysaccharide iron complex. Result showed high iron(27.4%) of it by atomic spectrophotometry. And in the 850cm-1 and 650cm-1 Office had characteristic peaks by infrared spectroscopy, indicating its iron core wasβ- FeOOH. And also, Ulva pertusa polysaccharide iron complex possess effitive antiradiation activity,compared to the polysaccharide of Ulva pertusa, and the difference of them were not significant.2-DE study of spleen in mice radiation experiments fed polysaccharide and polysaccharide iron (Ⅲ) show that,compared to control group , 2 proteins are 2-fold up-regulated and 13 2-fold down-regulated in irradiation control group, and compared to irradiation control group , there are 14 2-fold up-regulated proteins , 5 2-fold down-regulated in polysaccharide group and 12 2-fold up-regulated proteins,8 2-fold down-regulated in polysaccharide iron group including 78kDa glucose regulated protein, prolyl hydroxylase beta, subunit calreticulin precursor, micro- tubulinβ5 and vimentin which all were associated with immune and injury.更多还原