摘要
目的探讨lasI/rhlI基因缺失和阿奇霉素对铜绿假单胞菌(PA)毒力基因表达的影响。方法将野生型及lasI/rhlI基因缺失的PA菌株(PA-ΔlasI,PA-ΔrhlI,PA-ΔlasIrhlI)培养6h,同时以2μg/mL的阿奇霉素处理上述菌株6h,采用荧光定量PCR技术观察lasI/rhlI基因缺失以及阿奇霉素对铜绿假单胞菌QS系统及其毒力基因表达的影响。结果缺失lasI基因后,其下游毒力基因lasA、aprX、rhlA、rhlB、phnA及phnB的表达水平均下调(P<0.05,P<0.01),而上述毒力基因在rhlI基因缺失后表达水平则无明显变化。阿奇霉素处理PA后,lasI基因的表达水平明显下降(P<0.01),而lasI下游毒力基因的表达水平明显增高(P<0.01,P<0.001),QS系统抑制剂qscR的表达水平也显著上调(P<0.001)。结论lasI基因缺失可以明显抑制铜绿假单胞菌QS系统毒力基因的表达,阿奇霉素可能通过抑制qscR而促进毒力基因的表达。
Objective To investigate the effect of lasI/rhlI gene deletion and azithromycin on the expres-sion of virulence gene of Pseudomonas aeruginosa. Methods PA strains of wild type and lasI/rhlI gene deletion(PA-ΔlasI,PA-ΔrhlI,PA-ΔlasIrhlI)were cultured for 6 hours,and the four strains were treated with 2 μg/m Lazithromycin for 6 hours. The effects of lasI/rhlI gene deletion and azithromycin on the QS system and the expres-sion of virulence genes in Pseudomonas aeruginosa were observed by fluorescence quantitative PCR. Results Afterdeletion of lasI gene,the expression of las A,apr X,rhl A,rhl B,phn A and phn B were all down-regulated(P <0.05,P < 0.01),while the expression of those genes did not change significantly after the deletion of the rhlI gene.After treatment with azithromycin,the expression of lasI gene was significantly decreased(P < 0.01),while the ex-pression level of virulence gene downstream of lasI was significantly higher(P < 0.01,P < 0.001),and the expres-sion level of QS system inhibitor qsc R was also increased(P < 0.001). Conclusion The deletion of lasI gene maysignificantly inhibit the expression of virulence genes of QS system in Pseudomonas aeruginosa,and azithromycinmay promote the expression of virulence genes by inhibiting qsc R.
引文
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