A preliminary study on siHybrids technique on inhibiting the efflux pump gene mexB of Pseudomonas aeruginosa in vitro
10.3760/cma.j.issn.0254-5101.2011.08.010
- VernacularTitle:siHybrids技术沉默铜绿假单胞菌外排泵 mexB基因体外效应的初步研究
- Author:
Yan MAO
;
Jia CHEN
;
Dong XU
;
Mingyou XING
;
Lili WANG
;
Xuhua XIE
;
Fengyun GONG
;
Chao XIA
;
Aixia SHEN
;
Ying SONG
- Publication Type:Journal Article
- Keywords:
siHybrids technique;
RNA interference;
Pseudomonas aeruginosa;
Effiux pump;
Real-time PCR
- From:
Chinese Journal of Microbiology and Immunology
2011;31(8):707-712
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect and mechanism of siHybrids technique on inhibiring the efflux pump gene mexB of Pseudomonas aeruginosa PAO1 in vitro. MethodsTargeting the efflux pump gene mexB of Pseudomonas aeruginosa PAO1 ,we designed and synthesized three siHybrids molecule and one negative scamble siHybrids molecule. Pseudomonas aeruginosa PAO1 were intervened by the siHybrids molecules in 50 nmol/L, respectively. And the experiments were made of control groups[blank and scamble (sc) -001]and intervened groups[siHybrids( si ) -001, siHybrids( si ) -002 and siHybrids(si) -003]of Pseudomonas aeruginosa PAO1. The targeting efflux pump gene mexB mRNA expressions of Pseudomonas aeruginosa PAO1, including all groups, were measured by real-time PCR in 12 h and 24 h after interference in vitro. Further, the minimal inhibitory concentration (MIC) of chlormycetinCP, erythrocin( EM ), levofloxacin ( L-OFLX), ceftazidime ( CAZ), meropenem (MER) to those groups were detected by using Mueller-Hinton broth dilution before and after interference. ResultsThe relative mexB mRNA amounts of Pseudomonas aeruginosa PAO1 intervened by different siHybrids were not much more different from each other after 12 h,but the expression of mexB mRNA of the intervened group ( si-001 ,si-002 ,si-003 ) was much lower than control groups after interference for 24 h. The relative mexB mRNA amounts, comparing 12 h with 24 h, we would find the blank control and negative control submit escalating trend. And the intervened control ,three different siHybrids were all with a downward tendency. However, in presence of 50 nmol/L siHybrids, the minimal inhibitory concentration(MIC) of CP, EM, L-OFLX, CAZ, MER to those controls were not much more different before and after interference. Conclusion On level of the mRNA expressions, siHybrids could inhibit the efflux pump gene mexB of Pseudomonas aeruginosa PAO1 in vitro, and within 24 h would be able to function effectively. Further, the effect was time-dependent.
