siHybrids对耐碳青霉烯类抗生素肺炎克雷伯菌kpc-2基因表达水平的影响  

Effect of short interfering hybrids on the expression of kpc-2 gene expression in carbapenem-resistant Klebsiella pneumoniae

作　　者：范芳华[1] 吴国友[1] 周筱琼[1] 王芳[1] 万晓晨[1] 

机构地区：[1]浙江医院检验科细菌室,浙江杭州310013

出　　处：《中华全科医学》2016年第6期1017-1018,1029,共3页Chinese Journal of General Practice

基　　金：浙江省医药卫生科技计划项目(2015KYA010)

摘　　要：目的研究si Hybrids对耐碳青霉烯类抗生素肺炎克雷伯菌kpc-2基因表达水平及其对美罗培南的抑菌效果的影响,探讨si Hybrids的作用机制。方法将48株携带有kpc-2基因的耐碳青霉烯类抗生素肺炎克雷伯菌株分成对照组(不加si Hybrid)、低浓度组(si Hybrids为0.125μg/ml)、高浓度组(si Hybrids为8.0μg/ml)。分别检测3组12、24、48 h后3个时间点的mRNA的量。另外再分别取3组菌液进行美罗培南药敏试验,观察3组间抑菌圈直径的大小有无差异。结果与对照组相比较,si Hybrids干预后kpc-2基因的mRNA表达量显著下降(P<0.05),高浓度组kpc-2基因的mRNA表达量显著低于低浓度组(P<0.05),si Hybrids的浓度与kpc-2基因的mRNA表达量呈负相关(r=-0.782,P=0.014);kpc-2基因的mRNA表达量随着si Hybrids干预时间的延长而显著下降(P<0.05)。si Hybrids干预能显著增大美罗培南抑菌圈的直径(P<0.05),不同浓度的si Hybrids对美罗培南抑菌圈直径的增大幅度不同,高浓度组美罗培南抑菌圈的直径显著大于低浓度组(P<0.05),si Hybrids的浓度与美罗培南抑菌圈直径的大小呈正相关(r=0.882,P=0.024)。结论 si Hybrids能降低耐碳青霉烯类抗生素肺炎克雷伯菌kpc-2基因的表达水平,能提高美罗培南的抑菌效果,其作用机制是si Hybrids分子特异性沉默了肺炎克雷伯菌的kpc-2基因。Objective To explore the influence of short interfering hybrids（ Sihybrids） on the expression level of kpc-2gene and the antibacterial effects of meropenem in carbapenem-resistant Klebsiella pneumoniae（ CRKP） and study the action mechanism of sihybrids. Methods Total 48 strains carrying kpc-2 gene resistant to carbapenems Klebsiella Klebsiella strains cultured per plant were divided into three copies,one without si Hybrids as control group,two other join si Hybrids as experimental group. The experimental group was divided into low concentration group（ si Hybrids interference dose for0. 125 μg / ml and high concentration group（ si Hybrids interfere with dose of 8. 0 μg / ml）. RT-PCR was performed in the three groups to detect the expression of the mRNA of kpc-2 gene in bacterial fluid taken at 12 h,24 h,and 48 h time points. The amount of mRNA at three time points were calculated and compared among the three groups to determine the effect of si Hybrids on the expression level of kpc-2 gene. The meropenem susceptibility tests were performed to observe the antibacterial circle diameter and infer indirectly the effects of si Hybrids on the expression yield of kpc-2. Results Compared with the control group,the mRNA expression of kpc-2 gene was significantly decreased after the intervention of siHybrids（ P 〈 0. 05）. The mRNA expression of kpc-2 gene in high concentration group was significantly lower than that in low concentration group（ P 〈 0. 05）; the concentration of si Hybrids was negatively correlated with the mRNA expression of kpc-2 gene（ r =- 0. 782,P = 0. 014）; the mRNA expression of kpc-2 gene decreased significantly（ P 〈 0. 05） with the prolongation of si Hybrids intervention time. Si Hybrids intervention can significantly increase the diameter of inhibition zone of meropenem（ P 〈 0. 05）,different concentrations of si Hybrids of meropenem antibacterial circle diameter increased dramatically different,group of high concentration of meropenem antibacterial circle dia

关 键 词：siHybrids 耐药 碳青霉烯类抗生素 肺炎克雷伯菌 kpc-2基因 碳青霉烯酶kpc-2 mRNA 

分 类 号：R446.5[医药卫生—诊断学] R978.1[医药卫生—临床医学]