To illustrate dynamics of secretion and the peptides patterns of Rana dybowskii, the crude skin secretions were collected at different time when their secretions were released with a mild electrical stimulation. The results showed that the restoring amount of skin secretions increased in a characteristic sigmoid curve during 0-21 days after an electrical stimulation. The HPLC showed that the skin secretions collected at different time had the identical components, however the components restored asynchronously. Two quickly-restored components peaks were tested for their antibacterial activity against Escherichia coli ATCC25922 and Staphylococcus aureus ATCC25923 and showed strong antibacterial activity. The two components were identified by ESI-MS/MS and their amino acid sequences were determined. The results indicated that the recovery rates of skin secretion components in R. dybowskii were asynchronous and the antimicrobial peptides had precedence over other peptides. The immune defense strategy would ensure the survival of frogs in a complex environment.%东北林蛙皮肤分泌物排空后,在不同时间点收集其皮肤分泌物,分析皮肤分泌物及其中抗菌肽谱的动态恢复规律.结果表明:电刺激排空后的第0 ～21天时间范围内东北林蛙皮肤分泌物总量的恢复呈递增趋势；高效液相色谱显示不同时间收集的皮肤分泌物的组分差异不显著(P ＞0.05,n=5),但各组分恢复的时间并不同步；用大肠杆菌和金黄色葡萄球菌对恢复较快的2个组分峰成分进行抑菌试验,发现2组分均有较强的抑菌活性.用ESI-MS/MS对2组份进行质谱鉴定,确定其氨基酸序列.东北林蛙皮肤分泌物中抗微生物肽的恢复表达优先于其他多肽,该免疫防御策略可为东北林蛙在复杂环境中的生存提供保证.
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