黄粉虫幼虫抗肿瘤细胞K562抗菌肽的分离纯化  

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

作　　者：刘颜岗[1] 程璟侠[2] 赵瑞君[1] 樊宏英[1] 

机构地区：[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

出　　处：《中国媒介生物学及控制杂志》2009年第6期565-568,共4页Chinese Journal of Vector Biology and Control

基　　金：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)

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。Objective To isolate and purify the antimicrobial peptides with antitumor 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, centrifugalization, 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-bacterial substances which have anti-K562 activity in the Tenebrio molitor L.larvae, and more than one.

关 键 词：黄粉虫幼虫 抗菌肽 肿瘤细胞 K562 纯化 

分 类 号：R384[医药卫生—医学寄生虫学]