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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 antimicrobial peptides against K562 cells from the body of Tenebrio molitor. Methods Ultrasonic induced Tenebrio larvae to express a large number of antimicrobial peptides. After being ground, centrifuged, solid phase extracted, reversed-phase high performance liquid chromatography, the antibacterial peptides of the larvae of Tenebrio molitor were separated and purified, and the antibacterial activity against K562 was screened by MTT colorimetric method and light microscope observation. Peptide. Results Centrifugation supernatant was loaded on the solid phase extraction column and eluted with 10%, 30%, and 80% acetonitrile solution. Only 80% of the acetonitrile aqueous solution fractions were active (P<0.01). After purified by reversed-phase high-performance liquid chromatography, the active components were separated into five peak substances with anti-tumor activity. The activity of these five peaks was strong (P<0.01), and peaks 9 and 4 could be initially identified as antibacterial. Peptides, the other three kinds need further proof of the experiment. Conclusion There are more than one kind of antibacterial peptides and antibacterial substances against tumor cell K562 in the larvae of Tenebrio molitor.