大肠杆菌胁迫下东北林蛙皮肤抗菌肽分泌特性研究  

Secretary Characteristics of Antimicrobial Peptides from Skin of Rana dybowskii under Escherichia coli Challenge

作　　者：徐佳佳[1] 肖向红[1] 柴龙会[1] 苗慧敏[1] 杨淑静[1] 

机构地区：[1]东北林业大学野生动物资源学院

出　　处：《经济动物学报》2010年第2期102-106,共5页Journal of Economic Animal

基　　金：国家自然科学基金项目(30870301);黑龙江省自然科学基金重点项目(ZJN0604-02)

摘　　要：以东北林蛙（Rana dybowskii）为试验对象,采用大肠杆菌（Escherichia coli）作为环境胁迫因子,通过RP-HPLC分析、抑菌活性和蛋白浓度比较,观察其皮肤分泌物的特异性分泌,以期揭示两栖类动物抵御环境微生物时皮肤分泌抗菌肽的变化规律。研究发现,用E.coli胁迫1~18d后皮肤分泌物粗提物洗脱峰数量明显减少了61.54%~84.62%,对4个持续出现的8、9、11和14号洗脱峰的抑菌活性试验显示,8、9号峰对G-和G＋菌均表现较强的活性抑制,而11、14号峰仅对G＋菌有较强的抑菌活性。此外,E.coli胁迫后第7天组皮肤分泌物的洗脱峰数量、持续出现峰的峰面积值及蛋白浓度与对照组比均明显增加,第23天组均恢复至较高水平。研究结果表明：两栖类皮肤抗菌肽可能在参与机体先天免疫系统防御和启动后天免疫应答的关键环节中发挥重要的桥梁作用。To reveal the changes of amphibian skin secretions under microbial challenge,we chosen the brown frog（Rana dybowskii）as the test object and E.coli as the stress factor,which is one of the most common environmental microorganisms.The changes after challenge were analyzed through RP-HPLC,protein concentration and antibacterial activity comparison.The RP-HPLC results showed that the number of peaks of skin secretions after challenge reduced to 61.54%-84.62%,and the antibacterial activity analysis of 4 peaks（number 8,9,11 and 14）which continuously appeared showed that peak 8,9 had strong inhibition activity to G-and G＋bacteria,peak 11,14 had strong inhibition activity to G＋bacteria.In addition,contrast to the control group,the number of RP-HPLC peaks,peak area value of four continuously appeared peaks and protein concentration of the skin secretion were significantly increased at 7th day after challenge,and all returned to a higher level at 23rd day.The results indicated that the antimicrobial peptide of amphibian skin secretions may play an important role in the critical link between innate and adaptive immunity.

关 键 词：东北林蛙 抗菌肽 大肠杆菌 胁迫 反相高效液相色谱 

分 类 号：S865.9[农业科学—野生动物驯养]