亚抑菌浓度亚胺培南对MRSA体外增殖及毒力相关基因mRNA表达水平的影响
发布时间：2018-08-05 17:24
【摘要】：目的:探讨亚抑菌浓度亚胺培南对耐甲氧西林金黄色葡萄球菌(MRSA)生物学活性的影响,阐明碳青霉烯类抗生素对MRSA活性的抑制作用及其机制,为临床应用碳青霉烯类抗生素治疗MRSA感染提供依据。方法:选取5株ST239型MRSA临床分离株,采用1/10和1/2最小抑菌浓度(MIC)亚胺培南与其体外共培养1.5、6.0和12.0h,分为对照组(培养液中不加亚胺培南)、1/10MIC组(培养液中添加1/10MIC浓度亚胺培南)和1/2MIC组(培养液中添加1/2MIC浓度亚胺培南)。采用荧光定量PCR方法检测各组MRSA株毒力相关基因纤维黏连蛋白A(fnbA)、葡萄球菌蛋白A(spa)、α溶血素(hla)、白细胞毒素D(lek-D)和E(lek-E)、肠毒素A(sea)mRNA相对表达水平,分光光度法检测各组MRSA株体外增殖活性。结果:与对照组比较,体外共培养6.0和12.0h时,1/2MIC组MRSA株增殖活性明显降低(P0.01);培养1.5和6.0h时,6个MRSA毒力相关基因mRNA相对表达水平均明显低于对照组(P0.01);培养12h时,1/10MIC和1/2MIC浓度组MRSA株各毒力相关基因mRNA表达未能检测到。结论:亚抑菌浓度亚胺培南对ST239型MRSA多种毒力相关基因mRNA表达具有明显抑制效应,高浓度亚胺培南可抑制MRSA的体外增殖,提示亚胺培南对于重症MRSA感染患者具有潜在的应用价值。
[Abstract]:Objective: to investigate the effect of imipenem on the biological activity of methicillin-resistant Staphylococcus aureus (MRSA) and to elucidate the inhibitory effect of carbapenem antibiotics on MRSA activity and its mechanism. To provide the basis for clinical application of carbapenem antibiotics in the treatment of MRSA infection. Methods: five clinical isolates of ST239 type MRSA were selected. 1 / 10 and 1 / 2 minimum inhibitory concentrations of (MIC) imipenem were co-cultured with it for 1.5 and 12.0h in vitro, and were divided into control group (without imipenem in culture medium) and imipenem at 1 / 10 1/10MIC concentration in culture medium and 1/2MIC group with 1/2MIC concentration in culture medium. The virulence related gene A (fnbA), staphylococcal protein A (spa), 伪 -hemolysin (hla), leukocytotoxin D (lek-D) and E (lek-E), enterotoxin A (sea) mRNA were detected by fluorescence quantitative PCR, and the proliferative activity of MRSA strain in vitro was detected by spectrophotometry. Results: compared with the control group, The proliferative activity of 1 / 2 MIC group was significantly decreased (P0.01), the relative expression level of 6 MRSA virulence related genes mRNA was significantly lower than that of the control group (P0.01) at 1.5 and 6.0 h, and the virulence of 1 / 10 MIC group was significantly lower than that of the control group (P0.01) at 12 h, and the virulence of MRSA strain in 1 / 10 MIC group and 1/2MIC concentration group was significantly lower than that in the control group at 12 h after incubation (P0.01). Gene mRNA expression was not detected. Conclusion: imipenem has obvious inhibitory effect on the expression of ST239 type MRSA virulence related gene mRNA, and high concentration of imipenem can inhibit the proliferation of MRSA in vitro. It suggests that imipenem has potential application value in patients with severe MRSA infection.
【作者单位】： 内蒙古医科大学附属医院检验科;内蒙古医科大学科技处;内蒙古医科大学微生物与免疫研究中心;
【基金】：国家自然科学基金资助课题(81260244,81660352) 内蒙古自治区科技厅科技计划资助课题(20140144);内蒙古自治区科技厅自然科学基金面上项目资助课题(2016MS0829) 内蒙古自治区卫计委科研项目资助课题(201303061)
【分类号】：R446.5
本文编号：2166471
[Abstract]:Objective: to investigate the effect of imipenem on the biological activity of methicillin-resistant Staphylococcus aureus (MRSA) and to elucidate the inhibitory effect of carbapenem antibiotics on MRSA activity and its mechanism. To provide the basis for clinical application of carbapenem antibiotics in the treatment of MRSA infection. Methods: five clinical isolates of ST239 type MRSA were selected. 1 / 10 and 1 / 2 minimum inhibitory concentrations of (MIC) imipenem were co-cultured with it for 1.5 and 12.0h in vitro, and were divided into control group (without imipenem in culture medium) and imipenem at 1 / 10 1/10MIC concentration in culture medium and 1/2MIC group with 1/2MIC concentration in culture medium. The virulence related gene A (fnbA), staphylococcal protein A (spa), 伪 -hemolysin (hla), leukocytotoxin D (lek-D) and E (lek-E), enterotoxin A (sea) mRNA were detected by fluorescence quantitative PCR, and the proliferative activity of MRSA strain in vitro was detected by spectrophotometry. Results: compared with the control group, The proliferative activity of 1 / 2 MIC group was significantly decreased (P0.01), the relative expression level of 6 MRSA virulence related genes mRNA was significantly lower than that of the control group (P0.01) at 1.5 and 6.0 h, and the virulence of 1 / 10 MIC group was significantly lower than that of the control group (P0.01) at 12 h, and the virulence of MRSA strain in 1 / 10 MIC group and 1/2MIC concentration group was significantly lower than that in the control group at 12 h after incubation (P0.01). Gene mRNA expression was not detected. Conclusion: imipenem has obvious inhibitory effect on the expression of ST239 type MRSA virulence related gene mRNA, and high concentration of imipenem can inhibit the proliferation of MRSA in vitro. It suggests that imipenem has potential application value in patients with severe MRSA infection.
【作者单位】： 内蒙古医科大学附属医院检验科;内蒙古医科大学科技处;内蒙古医科大学微生物与免疫研究中心;
【基金】：国家自然科学基金资助课题(81260244,81660352) 内蒙古自治区科技厅科技计划资助课题(20140144);内蒙古自治区科技厅自然科学基金面上项目资助课题(2016MS0829) 内蒙古自治区卫计委科研项目资助课题(201303061)
【分类号】：R446.5
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