复合酶解制备海蜇抗肿瘤肽活性研究【毕业论文】.docVIP

本科毕业论文 （20 届） 复合酶解制备海蜇抗肿瘤肽活性研究 专业：药学 目录1 前言 3 2 实验材料与仪器 4 2.1材料与试剂 4 2.2主要仪器 4 3实验方法 4 3.1海蜇活性肽的制备 4 3.2分离、纯化海蜇抗肿瘤活性肽 5 3.3海蜇抗肿瘤肽的活性测定 5 4实验结果及讨论 6 4.1 分离纯化 6 4.2海蜇初 6 4.3 海蜇复合酶解液的抗肿瘤活性测定 8 5小结 11 [参考文献] 12  [摘要] 目的：从海蜇( Rhopilem aesculentum)中分离纯化活性肽，并研究其抗肿瘤活性。方法：海蜇经碱性蛋白酶初酶解、胰蛋白酶二次酶解后得复合酶解液，将酶解液、复合酶解液分别离心，收集上清液得海蜇活性肽粗品。粗品经超滤、SephadexG-15凝胶层析、冷冻干燥纯化得海蜇抗肿瘤活性肽，通过MTT法测定了初酶解产物对人前列腺癌细胞、肺癌细胞体外增殖的影响，以及复合酶解产物对人前列腺癌细胞、肺癌细胞、胃癌细胞体外增殖的影响。结果：从海蜇中分离纯化得到多种活性肽，MTT法表明了海蜇初酶解产物对肿瘤细胞的增殖有较强的抑制作用，复合酶解产物对肿瘤细胞具有抑制作用并有明显的量效依赖关系。结论：复合酶解制备的活性肽对肿瘤细胞有抑制作用。 [关键词] 海蜇；复合酶解；抗肿瘤；活性肽 Study on anti-tumor activity of the peptide Purified From jellyfish by composite enzymatic hydrolysis [Abstract] Objective: To isolate and purify the peptide from the jellyfish (Rhopilem aesculentum), and investigated its anti-tumor activity. Methods: First, jellyfish was hydrolysed by the alkaline protease enzyme and trypsin, we can get the enzyme solution, and after centrifugal the two kinds of enzyme solution, jellyfish crude peptides can be got by collectting the supernatant. Then, crude product was purified by ultrafiltration, SephadexG-15 gel chromatography, and freeze-drying and would to be the peptide with anti-tumor activity. Last, by MTT method, the enzymed solution’s influence on the proliferation in vitro of human prostate cancer, lung cancer cells was determined, as well as complex hydrolysate’s effect on proliferation of human prostate cancer, lung cancer cells and gastric cancer cells in vitro was determined, too. Results: From the jellyfish a variety of active peptides can be isolated and purified,and MTT method showed that the enzyme products’ effect on tumor cell proliferation was strongly inhibited and the compound hydrolyzate can inhibit the tumor cells with a significant dose dependent. Conclusion: The activie peptide prepared by composite enzymatic hydrolysis inhibited tumor cells. [Key words] jellyfish；composite enzymatic hydrolysis; anti-tumor；active peptides 1 前言 恶性肿瘤是严重危害人类健康的重大疾病，已成为我国人口死亡的最