作 者：
董亚萍;冯东岳;孙晶;张小明;胡鲲;杨先乐
单 位：
国家水生动物病原库/水产科学国家级实验教学示范中心/水产动物遗传育种中心上海市协同创新中心/上海海洋大学;全国水产技术推广总站
关键词：
连翘酯苷A;嗜水气单胞菌;恩诺沙星;耐药性;耐药节结化细胞分化家族(RND)
摘 要：
【目的】明确连翘有效成分延缓嗜水气单胞菌(Aeromonas hydrophila)对恩诺沙星的耐药性及其对药物主动外排作用中耐药节结化细胞分化家族(RND)相关基因表达的影响,为明确连翘延缓氟喹诺酮类药物耐药性的有效成分及揭示其延缓机制提供参考依据。【方法】观测连翘提取液及其与恩诺沙星联合作用对嗜水气单胞菌耐药菌株生长的影响,测定连翘有效成分(连翘苷、连翘酯苷A和连翘酯苷B)作用于嗜水气单胞菌前后的最小抑菌浓度(MIC),并利用实时荧光定量PCR检测连翘脂苷A作用于嗜水气单胞菌耐药菌株后RND相关基因(aheA、aheB和aheC)的表达变化。【结果】连翘提取液与64μg/mL恩诺沙星联合作用下,连翘提取液对嗜水气单胞菌耐药菌株的MIC为3.12μg/mL;在MIC和2MIC的条件下,连翘提取液与恩诺沙星联合作用能有效抑制嗜水气单胞菌耐药菌株的生长,24 h内未见明显的生长趋势。连翘的3种有效成分对嗜水气单胞菌耐药菌株均表现出抑制作用,在1.00μg/mL浓度条件下,连翘脂苷A的整体抑制效果较连翘苷和连翘脂苷B的效果更好。以1.00μg/mL连翘脂苷A作用于嗜水气单胞菌耐药菌株后,aheA、aheB和aheC基因均呈不同程度的下调表达,其中aheA基因的相对表达量较连翘脂苷A作用前显著下降(P<0.05)。【结论】连翘提取液与适宜浓度的恩诺沙星联合使用对嗜水气单胞菌有明显抑制效果,同时能延缓嗜水气单胞菌的耐药性,其中又以连翘脂苷A对RND主动外排系统的抑制作用最明显。
译 名：
Delaying effect of forsythiaside A on Aeromonas hydrophila resistance to enrofloxacin and its exocytosis
作 者：
DONG Ya-ping;FENG Dong-yue;SUN Jing;ZHANG Xiao-ming;HU Kun;YANG Xian-le;National Pathogen Collection Center for Aquatic Animals/National Demonstration Center for Experimental Fisheries Science Education/Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding/Shanghai Ocean University;National Fisheries Technical Extension Center;
单 位：
DONG Ya-ping%FENG Dong-yue%SUN Jing%ZHANG Xiao-ming%HU Kun%YANG Xian-le%National Pathogen Collection Center for Aquatic Animals/National Demonstration Center for Experimental Fisheries Science Education/Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding/Shanghai Ocean University%National Fisheries Technical Extension Center
关键词：
forsythiaside A;;Aeromonas hydrophila;;enrofloxacin;;antibiotic resistance;;resistant nodule cell differentiation family(RND)
摘 要：
【Objective】The delaying effect of active ingredients of Forsythia on the antibiotic resistance of Aeromonas hydrophila to enrofloxacin was explored. And the effects of active ingredients of Forsythia on related gene expression of resistant nodule cell differentiation family(RND)in active exocytosis of drugs were investigated in order to provide referential basis for discovering the effective ingredients of Forsythia to delay the antibiotic resistance of fluoroquinolone and revealing its delay mechanism.【Method】The effects of Forsythia extract and its combination with enrofloxacin on the growth situation of A. hydrophila antibiotic-resistant strain were observed. The minimum inhibitory concentration(MIC)of three active ingredients of Forsythia(forsythin,forsythiaside A and forsythiaside B)were determined before and after their treatments on A. hydrophila. Real-time fluorescence quantification PCR was used to detect the expression variation of three RND related genes(aheA,aheB,and aheC)after A. hydrophila antibiotic-resistant strain was treated by forsythiaside A.【Result】The MIC of Forsythia extract against A. hydrophila antibiotic-resistant strain was 3.12 μg/mL under the combination of Forsythia extract and 64 μg/mL enrofloxacin. The combination of Forsythia extract and enrofloxacin could effectively inhibit the growth of A. hydrophila antibiotic-resistant strain under the conditions of MIC and 2MIC. And therewas no obvious growth tendency within 24 h. The three active ingredients of Forsythia showed inhibitory effects on A. hydrophila antibiotic-resistant strain. At a concentration of 1.00 μg/mL,the inhibitory effect of forsythiaside A was better than that of forsythin and forsythiaside B. After treatment with 1.00 μg/mL forsythiaside A on A. hydrophila antibiotic-resistant strain,the aheA,aheB and aheC genes all showed down-regulated expression to different degrees. And the relative expression of aheA gene was significantly decreased(P<0.05)compared with that before forsythiaside A treatment.【Conclusion】The combination of Forsythia extract and enrofloxacin at a suitable concentration has obvious inhibitory effects on A. hydrophila and can delay theantibiotic resistance of A. hydrophila. Forsythiaside A has the largest inhibitory effect on the RND active exocytosis system.
