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目的诱导家蝇产生抗菌活性肽,鉴定其抗菌活性。方法家蝇经针刺损伤感染大肠埃希菌,免疫诱导24h后制成组织匀浆,经低温盐析、超速离心、Sephadex G50层析、RP-HPLC等技术分离纯化抗菌肽。K-B法和稀释法测定抗菌活性。结果家蝇体内含有抗菌活性成分,家蝇经损伤免疫诱导后分泌抗菌肽的量明显增加,经分离纯化得到了一个色谱纯的抗菌活性肽。该抗菌肽对标准菌株大肠埃希菌(ATCC25922)、铜绿假单胞菌(ATCC27853)和金黄色葡萄球菌(ATCC25923)均有抗菌活性,其MIC分别为2.4μmol/L、3.6μmol/L和4.8μmol/L。结论免疫诱导可促进家蝇抗菌肽的分泌,分泌的抗菌肽有广谱抗菌活性,对革兰阴性菌和革兰阳性菌均有抑制作用。
Objective To induce Musca domestica to produce antibacterial peptides and identify its antibacterial activity. Methods Escherichia coli was infected by acupuncture in housefly. Immunohistochemistry was used to prepare tissue homogenate after 24 h. The antimicrobial peptides were isolated and purified by cryogenic saltation, ultracentrifugation, Sephadex G50 chromatography and RP-HPLC. K-B method and dilution method for the determination of antibacterial activity. Results Musca domestica contained antibacterial active ingredients. The quantity of antibacterial peptides secreted by Musca domestica was significantly increased after induced by injury, and a chromatographic pure antibacterial peptide was obtained after isolation and purification. The antibacterial peptides have antibacterial activity against standard strains of Escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC27853) and Staphylococcus aureus (ATCC25923) with MICs of 2.4 μmol / L, 3.6 μmol / L and 4.8 μmol / L. Conclusion Immune induction can promote the secretion and secretion of antibacterial peptides of Musca domestica with broad-spectrum antimicrobial activity and inhibit both Gram-negative and Gram-positive bacteria.