绵羊脾脏转移因子的制备和理化性质检测及对小鼠免疫功能影响

发布时间：2018-07-18 10:37

【摘要】：目的：本研究以绵羊脾脏为制备转移因子(TF)的原料,进行绵羊脾脏转移因子(S-TF)制备方法的探究,明确S-TF的制备原理及制备方法。首先,对S-TF进行初步的理化性质分析；其次进行动物试验,探究S-TF是否会对动物器官产生致病变作用以及是否具有提高动物免疫功能的作用。方法：本研究以相关文献资料为依据,采取冻融-透析的方法进行S-TF的制备。制备步骤经过反复摸索确定后,先后开展了S-TF的制备、初步的理化性质分析、实验动物安全性评价和小鼠免疫功能影响的探究。理化性质分析包括：外观性状、pH值、蛋白质反应、多肽含量的测定、核糖含量的测定、核苷酸与多肽的比值(OD260/OD280)的测定、氨基酸种类及含量的测定、矿物元素含量的测定、无菌检验等。动物试验分两部分：第一部分是将粗提的S-TF给小鼠灌服15d,首先眼球采血进行血常规检测,其次测定脾脏指数,再次取心脏、肺脏、肝脏、脾脏、肾脏、胃、十二指肠、空肠、回肠、盲肠、结肠、直肠等器官制作病理切片并进行病理组织学检查；第二部分是进行为期19d的小鼠腹腔巨噬细胞吞噬鸡红细胞试验,测定吞噬百分率和吞噬指数,通过上述两部分试验：检验S-TF是否具有提高免疫功能的作用。结果：通过对S-TF制备方法的反复摸索,已经掌握了基本制备原理和方法。通过初步的理化性质分析,各项测定结果均符合TF的国家标准。动物试验的结果显示：试验组小鼠的白细胞数,特别是中性粒细胞数目显著升高；光学显微镜下观察小鼠主要组织器官无病理变化；脾脏指数无显著差异；S-TF可以提高小鼠腹腔巨噬细胞的吞噬百分率和吞噬指数等结果,表明S-TF具有提高小鼠的免疫功能的作用。
[Abstract]:Objective: to investigate the preparation method of sheep spleen transfer factor (S-TF) using sheep spleen as the raw material to prepare transfer factor (TF), and to clarify the preparation principle and method of S-TF. First, the physical and chemical properties of S-TF were analyzed, and then the animal experiments were carried out to find out whether S-TF could cause pathological changes in animal organs and improve the immune function of animals. Methods: in this study, S-TF was prepared by freezing-thawing-dialysis. After repeated exploration, the preparation of S-TF, the preliminary analysis of physical and chemical properties, the safety evaluation of experimental animals and the influence of mouse immune function were carried out. Physical and chemical properties include: physical and chemical properties: pH value, protein reaction, determination of polypeptide content, determination of ribosomal sugar content, determination of the ratio of nucleotide to peptide (OD260 / OD280), determination of amino acid type and content, determination of mineral element content. Aseptic examination, etc. The animal experiment was divided into two parts: in the first part, the crude S-TF was infused to mice for 15 days. First, blood was taken from eyeball for routine examination, then spleen index was measured, heart, lung, liver, spleen, kidney, stomach, duodenum, jejunum were taken again. Pathological sections of ileum, cecum, colon and rectum were made and histopathological examination was carried out. The second part was the phagocytosis of chicken erythrocytes by peritoneal macrophages of mice for 19 days, and the phagocytic percentage and phagocytosis index were measured. Through the above two parts of the experiment: to test whether S-TF has the function of improving immune function. Results: the basic preparation principle and method have been mastered through repeated exploration of the preparation method of S-TF. Through the preliminary analysis of physicochemical properties, the results are in accordance with TF national standard. The results of animal experiments showed that the number of white blood cells, especially neutrophils, was significantly increased in the experimental group, no pathological changes were observed in the main tissues and organs under optical microscope, and there was no significant difference in spleen index between the experimental group and the control group. S-TF could increase the phagocytic percentage and phagocytic index of mouse peritoneal macrophages, which indicated that S-TF could improve the immune function of mice.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.4

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