中国媒介生物学及控制杂志 ›› 2009, Vol. 20 ›› Issue (6): 565-568.

• 论著 • 上一篇    下一篇

黄粉虫幼虫抗肿瘤细胞K562抗菌肽的分离纯化

刘颜岗1,程璟侠2,赵瑞君1,樊宏英1   

  1. 1 山西医科大学寄生虫学教研室(太原 030001); 2 山西省疾病预防控制中心
  • 收稿日期:2009-07-13 出版日期:2009-12-20 发布日期:2009-12-20
  • 通讯作者: 赵瑞君, Email: 4935186aaa2272@sina.com
  • 作者简介:刘颜岗(1981-),男,河北邯郸人,硕士研究生,从事昆虫抗肿瘤细胞抗菌肽的筛选和纯化。
  • 基金资助:

    山西省高校科技研究开发项目(200613011);山西省科技攻关项目(2006031087-04);太原市科学技术发展计划项目(081049)

Isolation and purification of antibacterial peptides with anti?K562 activity from the Tenebrio molitor Linnaeus larvae 

 LIU Yan-Gang, CHENG Jing-Xia, ZHAO Rui-Jun, FAN Hong-Ying   

  1. Institute of Parasitology, Department of Parasitology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Received:2009-07-13 Online:2009-12-20 Published:2009-12-20
  • Contact: ZHAO Rui-jun, Email: 4935186aaa2272@sina.com

摘要:

 【摘要】 目的 从黄粉虫幼虫体内分离纯化具有抗肿瘤细胞K562的抗菌肽。方法 通过超声诱导黄粉虫幼虫大量表达抗菌肽。然后经过研磨、离心、固相萃取、反相高效液相色谱分离纯化黄粉虫幼虫抗菌肽,采用四甲基偶氮噻唑蓝(MTT)比色法和光镜观察法筛选对K562有杀伤作用的抗菌肽。结果 离心上清液上样固相萃取柱,经10%、30%、80%的乙腈水溶液洗脱,只有80%的乙腈水溶液分离组分有活性(P<0.01)。该活性组分经反相高效液相色谱纯化后分离出5个具有抗肿瘤活性的峰物质,这5个峰活性都较强(P<0.01),其中峰9、峰4能初步确定为抗菌肽,其他3种有待于进一步的实验证明。结论 黄粉虫幼虫体内存在抗肿瘤细胞K562的抗菌肽和抗菌物质,而且不止一种。

关键词: 黄粉虫幼虫, 抗菌肽, 肿瘤细胞, K562, 纯化

Abstract:

 【Abstract】 Objective To isolate and purify the antimicrobial peptides with anti-tumor cell K562 activity from the Tenebrio molitor Linnaeus larvae. Methods Antimicrobial peptides of Tenebrio molitor L.larvae induced by ultrasonic waves were isolated and purified by trituration,centrifuga1ization,solid phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP?HPLC). Then the antimicrobial peptides with anti-K562 activity were sieved by MTT colorimetric method and light microscope observation. Results Supernatant eluted with 10%, 30%, 80% acetonitrile (ACN) in aqueous solution by solid phase extraction, of which, only 80% was active (P<0.01). Five anti-tumor peaks appeared after purification by RP-HPLC, which all had strong activity (P<0.01). Only 9 and 4 peaks could initially identified as antimicrobial peptides, the others still need to be proved. Conclusion There are antimicrobial peptides and anti-ba

Key words: Tenebrio molitor Linnaeus larvae, Antimicrobial peptides, Tumor cells, K562, Purification

中图分类号: