人IIA型PLA2衍生肽分子CT17杀菌活性研究.docxVIP

人IIA型PLA2衍生肽分子CT17杀菌活性研究 淳于柳爽梁宁牛(通讯作者) (广西医科大学附属肿瘤医院药学部广西南宁530021) 【摘要】目的 研究和比较人IIA型磷脂酶A2(phospholipase A2, PLA2)衍生多肽 CT17对不同细菌在体外的杀菌效应。方法 根据人IIA型PLA2氨基酸顺序分别 设计合成肽分子CT17 (p51-67)o采用琼脂铺板计数法测定杀菌活性，将不同浓 度衍牛肽分别与革兰氏阳性菌(G+)金黄色葡萄球菌、枯草杆菌、屎肠球菌和 革兰氏阴性菌(G?)大肠杆菌、绿脓杆菌、鲍曼不动杆菌在37°C孵育2.5 h,然后铺 板并置于37°C恒温箱培养18?24h,记录每一琼脂板上的菌落数(CFU),并计算 多肽作用后的杀菌率。结果IIA型PLA2CT17对G+菌与G■菌均有较强的杀菌活性。 结论 衍牛自人II A型PLA2保守区域17个氨基酸残基的肽M17对G+菌和G■均 有较强的杀菌活性，这可能与M17肽分子更易于与细菌结合并发挥作用有关。 【关键词】人IIA型磷脂酶A2保守区域抗菌肽衍生肽 【中图分类号】R96 【文献标识码】A 【文章编号】2095-1752 (2013) 17-0128-02 [Abstract] Object To study a nd compare the bactericidal activity of the polypeptide CT17 which derives from human Group IIA phospholipase A2 (PLA2, phospholipase A2) in vitro. Methods Peptide CT17(p51-67) was synthesized according to the sequenee of human Group IIA PLA2 amino acid. Six species of bacterial, Gram positive(G+) Staphy lococcus aureus , Bacillus subtilis , Enterococcus faecium , Gram negative(G-) Escherichia coli, Acinetobacter baumannii, Bacillus pyocyaneus, were incubated with different concentration of polypeptides at 37°C for 2.5 hours in a water bath respectively. Then the reaction solution was diluted and agar plates were poured. After 18-24 hours incubation in the thermostated container at 37°C.The colony formed units were counted and the bactericidal rates were calculated? Results PLA2CT17 possessed potent bactericidal activity on the G+ bacteria and G- bacteria. Conclusions: Peptide CT17 derived from human II type A PLA2 highly con serve regi on possesses a strong bactericidal activity, this may be related to the CT7 peptide molecules more easily binding to bacteria? 【Key Word 】 Human Group IIA phospholipase A2 Conserved region Antibacterial peptid Derived peptide 临床上日益严重的的细菌耐药问题已经引起社会的广泛关注，无论在国 内还是国外，都在积极的面对这一问题。普遍认为，通过合理使用抗菌药物可以 延缓和控制耐药菌的产生，而对于耐药菌，则要依赖不断开发新型抗菌药⑴。 开发新型抗菌药的方式有多种，其中抗菌肽是研究热点之一。自然界中的动物, 昆虫甚至微生物都天然存在有抗菌肽，它们的分子大小可以从几个氨基酸残基到 几十个氨基酸残基，由于这些抗菌肽有着与目前临床上使用的抗菌药物完全不同 的杀菌机制，故被认为有可能成为成为替代常规抗菌药物用于对付耐药菌的新型 抗菌药[2]o IIA型磷脂酶A2 （phosp