牵牛子蛋白的提取及抗肿瘤活性评价

Extraction of protein from Pharbitidis semen and evaluation of anti-tumor activity

利用响应面法对牵牛子黑丑的总蛋白提取进行工艺优化并观察其抗肿瘤活性。采用单因素及响应面法,以牵牛子蛋白提取量为指标,考察料液比、温度和pH三个因素对蛋白质提取量的影响。在经过双酶解法、SephadexG-15凝胶层析后,利用MTT(噻唑蓝)实验观察多肽组分的抗肿瘤活性。响应面优化结果发现影响牵牛子总蛋白质提取的各因素主次顺序为料液比>温度>pH,且最佳提取工艺为:pH 8.0,料液比为1∶50,温度为57℃,牵牛子蛋白提取量为544.608±1.13 mg(n=3),与理论预测相对误差较小。MTT实验结果显示多肽组分GA和GB对人鳞状上皮舌癌细胞SCC9和SCC25活性有显著抑制作用,但对人乳腺癌细胞和肺癌人肺泡基底上皮细胞则没有明显抑制效果。研究结果显示:牵牛子蛋白提取工艺条件合理,其抗肿瘤活性的研究为后期治疗舌鳞状细胞癌临床药物的研发提供科学依据。

Objective:To optimize the extraction process of total protein from Pharbitidis semen and observe its anti-tumor activity by response surface methodology.Methods:Taking the extracted amount of Pharbitidis semen protein as the index,single factor and response surface methodology were used to investigate the effects of three factors,such as the ratio of material to liquid,temperature and pH.After double enzymolysis and Sephadex G-15 gel chromatography,the anti-tumor activity of polypeptide components was observed by MTT test.Results:Response surface optimization results showed that the main and secondary order of the factors affecting the extraction of total protein from Pharbitidis semen was the ratio of material to liquid>temperature>pH,and the optimal extraction process was pH 8.0,the ratio of material to liquid was 1∶50,the temperature was 57℃,and the extraction amount of Pharbitidis semen protein was 544.608±1.13 mg(n=3),which had a small relative error with the theoretical prediction.MTT test results showed that the polypeptide components GA and GB had significant inhibitory effects on the activities of human squamous epithelial tongue cancer cells SCC9 and SCC25,but had no significant inhibitory effect on human breast cancer cells and lung cancer human alveolar basal epithelial cells.Conclusions:The extraction process of Pharbitidis semen protein is reasonable,and the study of its anti-tumor activity provides scientific basis for the development of clinical drugs for the treatment of tongue squamous cell carcinoma.