舍曲林对副溶血弧菌的抑制作用

Inhibitory Effect of Sertraline on Vibrio Parahaemolyticus

近年来关于副溶血弧菌环境和海产品分离株出现抗生素耐药性的报道日益增多,亟需寻找替代传统抗菌剂抗菌的策略以控制副溶血弧菌的污染和感染。本研究通过最小抑菌浓度(MIC)和最低杀菌浓度(MBC)的测定、生长曲线的绘制,以及运动性实验和结晶紫染色实验,并利用透射电镜观察副溶血弧菌形态的变化,来评估以舍曲林为代表的5-羟色胺再摄取抑制剂(SSRIs)类药物对副溶血弧菌的抑制活性及抗生物被膜形成能力。同时,通过检测与分析舍曲林对副溶血弧菌毒力基因转录的影响,探究舍曲林对副溶血弧菌的减毒作用。结果显示,舍曲林对副溶血弧菌的MIC为32μg/mL,MBC为64μg/mL,能够损伤其细胞膜和细胞壁,MIC下导致其干瘪皱缩,MBC下引起其涨裂、胞内物质外泄;亚抑制浓度的舍曲林能显著抑制副溶血弧菌的泳动和群集运动,抑制率分别为88.6%和71.5%;舍曲林呈浓度依赖性地抑制副溶血弧菌的生物被膜形成,当舍曲林质量浓度分别为8、16、32和64μg/mL时,对副溶血弧菌生物被膜形成的抑制率分别为66.9%、79.5%、88.3%和89.3%;亚抑制浓度的舍曲林能显著抑制副溶血弧菌的毒力基因fliA、ompW和aphA的表达,抑制率分别为71.9%、88.7%和77.3%。实验结果表明,舍曲林对副溶血弧菌具有良好的抗菌活性与抗生物被膜形成能力,在不影响副溶血弧菌生长的浓度下,可降低其毒力基因的转录水平。

Recently,there have been increasing reports on the emergence of antibiotic resistance in environmental and seafood isolates of Vibrio parahaemolyticus.Therefore,it is urgent to find strategies to replace traditional antimicrobial agents for antibacterial control of Vibrio parahaemolyticus contamination and infection.In this study,the antibacterial activity and anti-biofilm formation ability of sertraline against Vibrio parahaemolyticus were evaluated through minimum inhibitory concentration(MIC)test,minimum bactericidal concentration(MBC)test,growth curve test,motivity test,and crystal violet staining test,and transmission electron microscope observation of the morphological changes of Vibrio parahaemolyticus.Meanwhile,the attenuating effect of sertraline on Vibrio parahaemolyticus was explored by detecting and analyzing the influence of sertraline on the transcription of virulence genes of Vibrio parahaemolyticus.The results indicate that the MIC of sertraline on Vibrio parahaemolyticus is 32μg/mL and the MBC is 64μg/mL,which are able to damage the cell membrane and cell wall of Vibrio parahaemolyticus.Furthermore,sertraline can lead to shrinkage and shriveling at MIC concentration,and rupture and leakage of intracellular substances at MBC concentration.Sertraline can significantly inhibit the swimming and swarm motion of Vibrio parahaemolyticus with the inhibition rates of 88.6%and 71.5%,respectively,at the subinhibitory concentration.The ability of sertraline to inhibit biofilm formation of Vibrio parahaemolyticus shows a linear correlation of sertraline’s concentration,and the inhibition rates of 8,16,32 and 64μg/mL sertraline on the biofilm formation of Vibrio parahaemolyticus ATCC17802 are 66.9%,79.5%,88.3%and 89.3%,respectively.Sertraline can significantly inhibit the expression of virulence genes fliA,ompW and aphA of Vibrio parahaemolyticus with inhibition rates of 71.9%,88.7%and 77.3%at subinhibitory concentration,respectively.The results show that sertraline has a good antibacterial activity and anti-biofilm formation ability against Vibrio parahaemolyticus,and it can reduce the transcription level of virulence genes under the allowable concentration of plasma without affecting its growth.