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摘要：[目的]本实验旨在研究体外消化的地龙蛋白对小鼠脾淋巴细胞增殖的影响。[方法]配制一定浓度地龙蛋白体外消化产物的RPMI-1640培养基，取小鼠脾淋巴细胞进行体外原代培养，加入有ConA作为有丝分裂原刺激其增殖，使用3H-TDR掺入法对增殖产生的细胞进行同位素标记，通过检测体外培养后细胞中的放射活度来表示淋巴细胞的增殖程度。[结果]地龙蛋白处理组的放射活度高于ConA（+）组，随着给药浓度的增加，放射活度先降低后升高。在同等浓度下，地龙蛋白处理组的dpm值比脱脂奶粉处理组的dpm值高，且差异显著。[结论]体外消化地龙产物对小鼠脾淋巴细胞的增殖具有明显的促进作用，且该作用大于相同浓度下的脱脂奶粉的作用。38795
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毕业论文关键词：地龙蛋白；体外消化；小鼠脾淋巴细胞；原代细胞培养；3H-TDR掺入法
Effects of Earthworm Protein in Vitro Digestion on Proliferation of Mice Splenic Lymphocyte
Abstract:[Objective]The purpose of this study was to investigate the effect of earthworm protein in vitro on the proliferation of splenic lymphocytes in mice.[Methods]RPMI-1640 medium of certain concentration of earthworm protein in vitro digestion was prepared. Splenic lymphocytes were taken for primary culture in vitro. ConA was added as mitogen to stimulate its proliferation. The proliferation of cells was isotopically labeled by 3H-TDR incorporation, and the extent of lymphocyte proliferation was determined by detecting the radiation intensity in the cells cultured in vitro. [Results]The radiation intensity of the earthworm protein treatment group is higher than that of ConA (+) group, and the radiation intensity decreases first and then increases with the increase of the concentration of the drug. At the same concentration, the DPM value of the earthworm protein treatment group was higher than that of the skim milk treatment group, and the difference was significant between the DPM value and the dry milk powder treatment group.[Conclusion]The product of in vitro digestion of earthworm can obviously promote the proliferation of splenic lymphocyte in mice, and the effect is greater than that of defatted milk powder of the same concentration.
Key words: Earthworm Protein；Vitro Digestion；Mice Splenic Lymphocyte；Primary Cell Culture；3H-TDR Incorporation Method
目 录
摘要1
关键词1
Abstract1
Key words1
引言（或绪论） 2
1材料与方法 2
1.1材料 2
1.1.1实验材料 2
1.1.2主要试剂 2
1.1.3主要仪器 3
1.2方法 3
1.2.1体外消化地龙蛋白 3
1.2.1.1地龙的冻干处理 3
1.2.1.2提取蛋白质 3
1.2.1.3体外消化 3
1.2.1.4消化产物的提纯 3
1.2.2相关溶液的配制 3
1.2.3小鼠脾淋巴细胞的制备 4
1.2.4 3H-TDR掺入法培养林白细胞4
1.2.5细胞的收集与放射性的测定 4
1.2.6数据处理 4
2 结果与分析4
2.1地龙的脱脂与体外消化 4
2.1.1地龙的脱脂 4
2.1.2地龙蛋白的体外消化 5
2.2各组淋巴细胞放射活度（dpm）的结果5
2.3 ConA对小鼠脾淋巴细胞增殖的影响5
2.4体外消化地龙蛋白对小鼠脾淋巴细胞增殖的影响 6
2.4.1体外消化地龙蛋白对小鼠脾淋巴细胞的促进增殖作用 6
2.4.2 体外消化地龙蛋白对小鼠脾淋巴细胞增殖的影响趋势6
2.5体外消化地龙蛋白与脱脂奶粉对小鼠脾淋巴细胞增殖的影响差异 7
2.5.1体外消化脱脂奶粉对小鼠脾淋巴细胞增殖的影响 7
2.5.2体外消化脱脂奶粉对小鼠脾淋巴细胞增殖的影响趋势 7
2.3.2体外消化地龙蛋白与脱脂奶粉对小鼠脾淋巴细胞的影响差异 8
3讨论 9
3.1地龙蛋白及多肽的制备方法探究 9
3.2地龙体内活性物质对机体免疫机能的影响 9