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目的:用蛋白酶酶解乌贼墨,经纯化获得寡肽,并对其进行抗肿瘤活性研究。方法:通过胰蛋白酶酶解获得乌贼墨酶解物,将酶解液用超滤、G-25分子筛凝胶层析进行分离纯化,高效液相进行纯度检测。分离纯化得到的寡肽进行氨基酸序列检测,并用CCK-8检测寡肽对DU-145细胞增殖的影响和HE染色方法观察其对细胞形态的影响。结果:乌贼墨寡肽的N端氨基酸序列为:Gln-Pro-Lys,得率为1.82%。将寡肽进行抗肿瘤活性研究,发现寡肽对DU-145细胞有明显的增殖抑制效应,且有浓度依赖性。HE染色结果显示了寡肽使细胞形态发生不规则变化,失去原有的恶性表型。结论:用酶解方法制得的乌贼墨寡肽有明显的抗肿瘤活性。
OBJECTIVE: To purify sepia by protease and obtain oligopeptides by purification, and to study its anti-tumor activity. Methods: The squid protein hydrolyzate was obtained by enzymolysis of trypsin. The hydrolyzate was separated and purified by ultrafiltration and gel filtration on G-25 molecular sieve. The purity of squid was detected by HPLC. The oligopeptides were isolated and purified to detect the amino acid sequence. The effects of oligopeptides on the proliferation of DU-145 cells were detected by CCK-8 and the cell morphology was observed by HE staining. Results: The N-terminal amino acid sequence of sepia oligopeptide was Gln-Pro-Lys with a yield of 1.82%. The oligopeptide antitumor activity study found oligopeptide DU-145 cells had a significant inhibitory effect on proliferation, and a concentration-dependent. The results of HE staining showed that the oligopeptide changed the cell morphology irregularly and lost the original malignant phenotype. Conclusion: The sepia oligopeptide obtained by enzymolysis has obvious antitumor activity.