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

doi:10.13481/j.1671-587x.20170304

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (03): 479-484.doi: 10.13481/j.1671-587x.20170304

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

王俊瑞¹, 段美庆¹, 额尔德木图², 孙鹏³, 魏常梅¹, 韩艳秋¹

1. 内蒙古医科大学附属医院检验科, 内蒙古呼和浩特 010050;
2. 内蒙古医科大学科技处, 内蒙古呼和浩特 010110;
3. 内蒙古医科大学微生物与免疫研究中心, 内蒙古呼和浩特 010059

收稿日期:2016-09-07 出版日期:2017-05-28 发布日期:2017-06-01
通讯作者: 韩艳秋,教授,主任医师,硕士研究生导师(Tel:0471-3451315,E-mail:qyh1016@sina.com) E-mail:qyh1016@sina.com
作者简介:王俊瑞(1981-),男,内蒙古自治区呼和浩特市人,副主任医师,医学博士,主要从事病原菌致病机制方面的研究。
基金资助:
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）；内蒙古自治区卫计委科研项目资助课题（201303061）；内蒙古自治区科技厅自然科学基金面上项目资助课题（2016MS0829）

Effects of sub-inhibitory concentration of imipenem on proliferation in vitro and mRNA expression levels of MRSA virulence related genes

WANG Junrui¹, DUAN Meiqing¹, Erdemutu², SUN Peng³, WEI Changmei¹, HAN Yanqiu¹

1. Clinical Laboratory, Affiliated Hospital, Inner Mongolia Medical University, Hohhot 010050, China;
2. Department of Science and Technolgy, Inner Mongolia Medical University, Hohhot 010110, China;
3. Microbiology and Immunity Research Centre, Inner Mongolia Medical University, Hohhot 010059, China

Received:2016-09-07 Online:2017-05-28 Published:2017-06-01

摘要/Abstract

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（P<0.01）；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（P<0.01）；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。

关键词: 耐甲氧西林金黄色葡萄球菌, 亚胺培南, 毒力, 细胞增殖, 亚抑菌浓度

Abstract: Objective: To explore the effect of sub-inhibitory concentration of imipenem on the bio-activities of methicillin resistant Staphylococcus aureus(MRSA)and illuminate the inhibitory effects of carbapenem antibioty on the activity of MRSA and their mechanisms,and to provide the basis for using the carbapenem antibiotics in the treatment of MRSA infection. Methods: Five ST239 type of MRSA clinical isolates were selected and were co-cultured with 1/10 and 1/2 minimal inhibitory concentration (MIC) of imipenem for 1.5,6.0 and 12.0 h,which were divided into control group(no imipenem),1/10MIC group(1/10MIC of imipenem),and 1/2MIC group(1/2MIC of imipenem).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes fibronectin A(fnbA),staphylococcal protein A(spa),α-hemolysin(Hla),leukocidin D(lek-D),leukocidin E(lek-E), and enterotoxin A in various groups;Spectrophotometry was used to detect the proliferation activity of MRSA strains in various groups in vitro. Results: After co-culture for 6.0 and 12.0 h,the proliferation activities of 5 trains of MRSA in 1/2MIC group in vitro were significantly decreased compared with control group (P<0.01).The relative mRNA expression levels of 6 virulence-related genes of 5 strains of MRSA in 1/10MIC and 1/2MIC groups were significantly decreased compared with control group(P<0.01).After co-culture for 12.0 h,the mRNA expressions of all the tested virulence-related genes were not found. Conclusion: The sub-inhibitory concentration of imipenem shows obviously inhibitory effect on the mRNA expressions of multiple virulence-related genes of ST 239 type of MRSA strains,and higher concentration of imipenem can suppress the proliferation of MRSA strains in vitro.Imipenem shows the potential value in the treatment of the severe MRSA infected patients.

Key words: virulence, sub-inhibitory concentration, methicillin resistant Staphylococcus aureus, cell proliferation, imipenem

中图分类号:

R378.1

王俊瑞, 段美庆, 额尔德木图, 孙鹏, 魏常梅, 韩艳秋. 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[J]. 吉林大学学报(医学版), 2017, 43(03): 479-484.

WANG Junrui, DUAN Meiqing, Erdemutu, SUN Peng, WEI Changmei, HAN Yanqiu. Effects of sub-inhibitory concentration of imipenem on proliferation in vitro and mRNA expression levels of MRSA virulence related genes[J]. Journal of Jilin University Medicine Edition, 2017, 43(03): 479-484.

参考文献

[1] Nelson RE,Stevens VW,Jones M,et al.Health care-associated methicillin-resistant Staphylococcus aureus infections increases the risk of postdischarge mortality[J].Am J Infect Control,2015,43(1):38-43.
[2] Cosgrove SE,Qi Y,Kaye KS,et al.The impact of methicillin resistance in Staphylococcus aureus bacteremia on patient outcomes:mortality,length of stay,and hospital charges[J].Infect Control Hosp Epidemiol,2005,26(2):166-174.
[3] Otto MP,Martin E,Badiou C,et al.Effects of subinhibitory concentrations of antibiotics on alpha-Toxin(hla) gene expression of methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolates[J].J Antimicrob Chemother,1998,68(7):1524-1532.
[4] Jo A,Ahn J.Phenotypic and genotypic characterisation of multiple antibiotic-resistant Staphylococcus aureus exposed to subinhibitory levels of oxacillin and levofloxacin[J].BMC Microbiol,2016,16(1):170.
[5] Jones M,Nielson C,Gupta K,et al.Collateral benefit of screening patients for methicillin-resistant Staphylococcus aureus at hospital admission:isolation of patients with multidrug-resistant gram-negative bacteria[J].Am J Infect Control,2015,43(1):31-34.
[6] Shang W,Hu Q,Yuan W,et al.Comparative fitness and determinants for the characteristic drug resistance of ST239-MRSA-Ⅲ-t030 and ST239-MRSA-Ⅲ-t037 strains isolated in China[J].Microb Drug Resist,2016,22(3):185-192.
[7] 王俊瑞,杜小莉,塔拉,等.甲氧西林耐药/敏感金黄色葡萄球菌基因分型和毒力基因检测[J].中国感染与化疗杂志,2015,15(1):70-75.
[8] Enright MC,Day NP,Davies CE,et al.Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus[J].J Clin Microbiol,2000,38(3):1008-1015.
[9] Rudkin JK,Laabei M,Edwards AM,et al.Oxacillin alters the toxin expression profile of community-associated methicillin-resistant Staphylococcus aureus[J].Antimicrob Agents Chemother,2014,58(2):1100-1107.
[10] Subrt N,Mesak LR,Davies J.Modulation of virulence gene expression by cell wall active antibiotics in Staphylococcus aureus[J].J Antimicrob Chemother,2011,66(5):979-984.
[11] Waters EM,Rudkin JK,Coughlan S,et al.Redeploying β-Lactam antibiotics as a novel antivirulence strategy for the treatment of methicillin-resistant Staphylococcus aureus infections[J].J Infect Dis,2017,215(1):80-87.
[12] Cheng VC,Li IW,Wu AK,et al.Effect of antibiotics on the bacterial load of meticillin-resistant Staphylococcus aureus colonisation in anterior nares[J].J Hosp Infect,2008,70(1):27-34.
[13] van Langevelde P,van Dissel JT,Ravensbergen E,et al.Antibiotic-induced release of lipoteichoic acid and peptidoglycan from Staphylococcus aureus:quantitative measurements and biological reactivities[J].Antimicrob Agents Chemother,1998,42(12):3073-3078.
[14] Santa Maria JP Jr,Sadaka A,Moussa SH,et al.Compound-gene interaction mapping reveals distinct roles for Staphylococcus aureus teichoic acids[J].Proc Natl Acad Sci USA,2014,111(34):12510-12515.
[15] Rudkin JK,Edwards AM,Bowden MG,et al.Methicillin resistance reduces the virulence of healthcare-associated methicillin-resistant Staphylococcus aureus by interfering with the agr quorum sensing system[J].J Infect Dis,2012,205(5):798-806.
[16] Singh R,Ray P.Quorum sensing-mediated regulation of staphylococcal virulence and antibiotic resistance[J].Future Microbiol,2014,9(5):669-681.
[17] Yan J,Liu Y,Gao Y,et al.RNAⅢ suppresses the expression of LtaS via acting as an antisense RNA in Staphylococcus aureus[J].J Basic Microbiol,2015,55(2):255-261.

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

Effects of sub-inhibitory concentration of imipenem on proliferation in vitro and mRNA expression levels of MRSA virulence related genes

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	朱德强, 陈学军. 白花丹素对肝癌索拉菲尼耐药细胞HepG2R增殖和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2018, 44(06): 1223-1229.
[2]	阎慧, 马淑飞, 段明华, 闫玉礼, 潘新, 李海清, 周长龙, 赵天倚. 京尼平对胃癌SGC 7901细胞体外增殖、侵袭能力和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2018, 44(05): 924-928.
[3]	邢树刚, 张丽红, 金明华, 李百慧, 武则馨, 房波, 李伟. 杨梅黄酮对小鼠脑胶质瘤GL261细胞增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(05): 955-961.
[4]	袁虎勤, 李强. 水飞蓟素对人胃癌SGC 7901细胞增殖、迁移和侵袭的抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(05): 988-993.
[5]	刘定坤, 杨楠, 金巨楼, 孙梦洁, 王倩, 吴佳, 刘志辉. 牙髓干细胞增殖、分化和迁移的研究进展[J]. 吉林大学学报(医学版), 2018, 44(05): 1100-1104.
[6]	王淼, 母润红, 李明成, 李洪杰. RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 753-758.
[7]	李鑫, 马云飞, 唐庚, 韦麒, 纪红池, 田嘉安, 申延男, 王志成. 线粒体靶向KillerRed诱导的ROS增强辐射对HeLa细胞的增殖抑制作用[J]. 吉林大学学报(医学版), 2018, 44(04): 718-723.
[8]	朱效慧, 孟琴, 冯世燕. TP53在不明原因复发性流产患者绒毛组织中的表达及其对人滋养层细胞生长的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 796-800.
[9]	吕鹏, 宁明杰, 邵玉, 张璐妮, 唐英, 王南博, 陈飞儿, 齐玲. 野黄芩苷对人脑胶质瘤U87细胞的增殖抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(03): 466-470.
[10]	杨文延, 刘强, 孙志娟, 杜利清, 徐畅, 王彦, 柳杨, 王芹. 褪黑素对人非小细胞肺癌H1299细胞辐射敏感性的影响[J]. 吉林大学学报(医学版), 2018, 44(03): 532-536.
[11]	周刘祥, 林琳, 胡鑫敏, 王晓春. 内分泌腺性血管内皮生长因子对结肠癌LoVo细胞增殖和迁移的促进作用[J]. 吉林大学学报(医学版), 2018, 44(03): 516-520.
[12]	上官梦原, 赵菁, 杨艳荣, 张佳悦, 陈俊宇, 赵淑华. 五味子脂A联合卡铂对人卵巢癌Skov3细胞增殖、迁移和侵袭的抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(02): 292-298.
[13]	张艳霞, 李跃辉, 张丽红, 张年萍, 闫燕艳, 杨小会, 冯湘玲. 沉默HOXA13基因对肝癌HepG₂和QGY-7703细胞恶性表型的影响[J]. 吉林大学学报(医学版), 2018, 44(02): 315-320.
[14]	毕然, 杜玉君, 魏伟, 王春喜. 穿心莲内酯对人肾细胞癌786-0细胞增殖、迁移和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(01): 18-23.
[15]	杨莉, 王雪雯, 周明, 张帆. As₂O₃联合γ分泌酶抑制剂MW167对人肝癌HepG₂/ADM细胞耐药性的影响[J]. 吉林大学学报(医学版), 2018, 44(01): 1-7.