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目的探讨氨溴索对铜绿假单胞菌生物膜(biofilm,BF)密度感应(quorum-sensing,QS)系统信号分子酰化高丝氨酸内酯(N-acyl-homoserinelactone,AHL)以及AHL合成酶编码基因表达的影响。方法通过平板培养法培养铜绿假单胞菌建立成熟的体外BF模型,用扫描电镜(scanning electron microscope,SEM)鉴定生物膜的形成情况。不同浓度氨溴索干预后,应用实时荧光定量PCR(real-time PCR)和AHL感应菌株根癌土壤杆菌(WCF47)分别检测其对铜绿假单胞菌AHL合成酶编码基因rhlI和lasI mRNA及AHL表达的影响。结果通过实时荧光定量PCR检测,结果显示氨溴索3.75mg/ml作用组基因lasI和rhlI与对照组相比表达量均显著减少(P<0.05)。氨溴索1.875mg/ml干预后,有同样趋势,但效应不如高浓度明显(P<0.05)。氨溴索3.75mg/ml作用后AHL的表达量较对照组明显减少(P<0.05),氨溴索1.875mg/ml作用后亦有减少(P<0.05)。结论氨溴索可以通过下调信号分子AHL合成酶编码基因以及AHL的表达来抑制BF。
OBJECTIVE: To investigate the effects of ambroxol on the biosynthesis of AHL and querum-sensing (QS) signaling molecules in biofilm (BF) biofilm (BF) Effects of gene expression. Methods Pseudomonas aeruginosa was cultured by plate culture to establish a mature in vitro BF model. The formation of biofilm was identified by scanning electron microscope (SEM). After different concentrations of ambroxol were interfered, real-time PCR and AHF-sensitive strain Agrobacterium tumefaciens (WCF47) were used to detect the expression of rhlI and lasI mRNA of AHL synthase gene and AHL The impact of expression. The results of real-time fluorescence quantitative PCR showed that the expression of lasI and rhlI in ambroxol 3.75mg / ml group was significantly decreased compared with the control group (P <0.05). Ambroxol 1.875mg / ml intervention, the same trend, but the effect was not as high concentration (P <0.05). Ambroxol 3.75mg / ml after AHL expression was significantly reduced compared with the control group (P <0.05), ambroxol 1.875mg / ml also decreased (P <0.05). Conclusion Ambroxol can inhibit BF by down-regulating the expression of AHL synthase gene and AHL.