| 刘 露,张月圆,李 开,王惠芸,王 冬,王灵芝.鸦胆子球蛋白酶解物对黑色素瘤细胞毒性研究[J].现代生物医学进展英文版,2018,(8):1447-1452. |
| 鸦胆子球蛋白酶解物对黑色素瘤细胞毒性研究 |
| The Cytotoxicity of Brucea Javanica Seeds Globulin Hydrolysates on Melanoma Cell |
| Received:December 08, 2017 Revised:December 30, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.08.009 |
| 中文关键词: 鸦胆子 球蛋白 提取工艺 抗肿瘤多肽 |
| 英文关键词: Brucea javancia Globulin Extraction process Antitumor peptides |
| 基金项目:北京中医药大学青年教师项目(2018-JYB-JS005） |
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| 中文摘要: |
| 摘要 目的：优化鸦胆子球蛋白提取工艺，采用酶解法获得多肽组分，并对其细胞毒性进行研究。方法：运用L9 (33) 表设计正交试验，筛选球蛋白提取最佳工艺参数，等电点法浓缩蛋白；采用不同蛋白酶水解球蛋白，经超滤（MWCO = 3 kDa）后收集滤过液，MTT法考察小分子量酶解物对鼠源黑色素瘤B16的细胞毒性。结果：球蛋白最佳工艺参数为：NaCl浓度5 %、提取时间1.5 h、固液比1: 12；胃蛋白酶为最优蛋白酶源，其水解产物（≤3 kDa）对B16细胞具有显著的细胞毒性，72 h时，IC50为1.08 μg?mL-1。结论：鸦胆子球蛋白源多肽组分具有明确的体外细胞毒性，有望进一步从中获得高活性抗肿瘤肽。 |
| 英文摘要: |
| ABSTRACT Objective: To optimize the extraction process parameter of globulin Brucea javanica seeds, obtain the polypeptide components by enzymatic hydrolysis, and evaluate its cytotoxicity. Methods: The orthogonal experiment of L9 (33) was firstly applied to obtain the optimal parameter for globulin extraction. The protein was then primarily purified by isoelectric precipitation. Different kinds of proteases were applied to hydrolyze B. javanica globulin followed by ultrafiltration with the molecular weight cut-off of 3 kDa to collect the permeate. MTT assay was then conducted to evaluate the cytotoxicity of low molecular weight hydrolysates (≤3kDa) on B16 cell line. Results: The optimal process parameters of the globulins extraction were as follows: 5 % of NaCl concentration, 1.5 h of extraction time and solid liquid ratio of 1:12; The hydrolysates (≤3kDa) of B. javanica globulin produced by pepsin showed significantly high growth inhibitory activity on B16 cells, with the IC50 value of 1.08 μg?mL-1 after 72 h incubation. Conclusion: The peptides of B. javanica globulin demonstrated specific cytotoxicity on melanoma cell line in vitro, suggesting antitumor peptides could be further obtained from B. javanica seeds. |
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