不同产地鸦胆子球蛋白源生物肽体外抗黑色素瘤活性研究
投稿时间:2019-04-09     点此下载全文
引用本文:王惠芸,陈佩瑶,欧阳雨晴,王冬,王灵芝.不同产地鸦胆子球蛋白源生物肽体外抗黑色素瘤活性研究[J].中国现代中药,2020,22(1):41-46
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作者中文名作者英文名单位中文名单位英文名E-Mail
王惠芸 WANG Hui-yun 北京中医药大学 生命科学学院,北京100029 Beijing University of Chinese Medicine,School of Life Sciences,Beijing 100029,China  
陈佩瑶 CHEN Pei-yao 北京中医药大学 生命科学学院,北京100029 Beijing University of Chinese Medicine,School of Life Sciences,Beijing 100029,China  
欧阳雨晴 OUYANG Yu-qing 北京中医药大学 生命科学学院,北京100029 Beijing University of Chinese Medicine,School of Life Sciences,Beijing 100029,China  
王冬 WANG Dong 北京中医药大学 中医学院,北京100029 Beijing University of Chinese Medicine,School of Chinese Medicine,Beijing 100029,China 王冬,硕士,助理研究员,研究方向:经方应用规律及机理研究;E-mail:wangdong_0701@126.com 
王灵芝 WANG Ling-zhi 北京中医药大学 生命科学学院,北京100029 Beijing University of Chinese Medicine,School of Life Sciences,Beijing 100029,China 王灵芝,博士,副研究员,研究方向:中药活性蛋白及多肽的功能研究;E-mail:wanglz@bucm.edu.cn 
基金项目:北京中医药大学青年教师项目(2018-JYB-JS005);国家自然科学基金项目(81872972)
中文摘要:目的:比较不同产地鸦胆子的主成分和球蛋白含量,进而分析各产地鸦胆子球蛋白酶解物对人源恶性黑色素瘤细胞A375的细胞毒性。方法:收集广东、广西、云南和福建4个产地鸦胆子,分别采用凯氏定氮法、旋光法、索氏提取法、烘干法、马弗炉法测定其蛋白质、淀粉、粗脂肪、水分和灰分含量。胃蛋白酶水解鸦胆子球蛋白,经超滤后收集滤过液,MTT法测定多肽组分对人源恶性黑色素瘤细胞A375的细胞毒性。结果:4个产地鸦胆子各成分存在显著差异,淀粉质量分数均在45%以上,粗脂肪质量分数为14.88%~22.10%,水分和灰分含量均符合《中华人民共和国药典》(2015版)规定;蛋白质质量分数以广西产地最高,为17.23%;福建产地最低,为12.61%。广西产地鸦胆子球蛋白含量为1.12 mg·(100 mg)-1,占总蛋白的6.52%,显著高于其他产地。4个产地鸦胆子球蛋白源小分子肽作用于A375细胞72 h后,最小半数抑制浓度(IC50)值分别为8.16、5.54、8.34、2.78 μg·mL-1。结论:不同产地对鸦胆子蛋白质含量存在一定影响,且球蛋白源生物肽均具有显著体外抗黑色瘤活性。
中文关键词:鸦胆子  球蛋白  黑色素瘤  细胞毒性  产地
 
In vitro Anti-melanoma Activity of Peptides Derived from Brucea javanica Globulin in Different Areas
Abstract:Objective:To compare the principal components and globulin content of Brucea javanica from different producing areas and then analyze the cytotoxicity of globulin hydrolysate to human melanoma cell line A375 cells.Methods:B.javanica seeds was collected from Guangdong,Guangxi,Yunnan and Fujian province.Kjeldahl method,polarimetry method,Soxhlet extraction method,drying method and muffle furnace method were applied to determinate the protein,starch,crude fat,water and ash content,respectively.Globulin was digested by pepsin and then the hydrolysate was fractioned by using ultrafiltration membrane with 3 kDa molecular weight cut-off to collect the filtered solution.Finally the cytotoxicity of polypeptides against A375 cells was evaluated by MTT method.Results: For the principal components,significant difference was detected among B.javanica species from four areas.The starch contents were all above 45% and the crude fat content ranged from 14.88% to 22.10%.The water and ash content were in accord with Pharmacopoeia of the People's Republic of China(ChP,2015 edition).The highest protein content of B.javanica was samples from Guangxi province accounting for 17.23%,while the lowest was 12.61% for the one from Fujian.The B.javanica globulin content from Guangxi was 1.12 mg·(100 mg)-1 accounting for 6.52% of total protein content,which was significantly higher than that of the other samples.The half maximal inhibitory concentration (IC50) values of small molecular peptides of globulin from Guangdong,Guangxi,Yunnan and Fujian regions on A375 cells were 8.16,5.54,8.34,2.78 μg·mL-1,respectively.Conclusion:Origins had certain effect on protein content of B. javanica and bioactive peptides of B.javanica globulin existed certain anti-melanoma activity in vitro.
keywords:Brucea javanica  globulin  melanoma  cytotoxicity  producing areas
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