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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 study the effect of si Hybrids on the expression of kpc-2 gene of carbapenem-resistant Klebsiella pneumoniae and its inhibitory effect on meropenem, and to explore the mechanism of si Hybrids. Methods 48 strains of Klebsiella pneumoniae resistant to carbapenem antibiotics carrying kpc-2 gene were divided into control group (without si Hybrid), low concentration group (0.125μg / ml for si Hybrids), high concentration group si Hybrids 8.0 [mu] g / ml). The mRNA levels of three groups after 12, 24, and 48 h were detected respectively. In addition, respectively, three groups of bacteria were taken Meropenem susceptibility test to observe the size of the inhibition zone diameter of the three groups there is no difference. Results Compared with the control group, the mRNA expression of kpc-2 gene in si-hybrids significantly decreased (P <0.05) and the mRNA expression of kpc-2 gene in high concentration group was significantly lower than that in low concentration group (P <0.05) si hybrids was negatively correlated with the mRNA expression of kpc-2 (r = -0.782, P = 0.014). The mRNA expression of kpc-2 was significantly decreased with the increase of si Hybrids intervention time (P <0.05 ). si Hybrids could significantly increase the diameter of meropenem inhibition zone (P <0.05). Different concentrations of si Hybrids increased the diameter of inhibition zone of meropenem, but the diameters of meropenem inhibition zone in high concentration group were significantly larger than that of meropenem (P <0.05). The concentration of si Hybrids was positively correlated with the size of the inhibition zone of meropenem (r = 0.882, P = 0.024). Conclusion si Hybrids can reduce the expression level of kpc-2 gene of carbapenem-resistant Klebsiella pneumoniae and enhance the antibacterial effect of meropenem. The mechanism is that si Hybrids molecule specifically silences Klebsiella pneumoniae Kpc-2 gene.