特异性阻断MecR1-MecI-MecA通路逆转耐甲氧西林金黄色葡萄球菌耐药性  

Reversal of antibiotic resistance in methicillin-resistant Staphylococcus aureus by blocking MecR1-MecI-MecA signal pathway

作　　者：孟静茹[1] 罗晓星[1] 扈本荃[2] 刘杰[1] 王海芳[1] 孟嘉[1] 

机构地区：[1]第四军医大学药学系药理学教研室,陕西西安710033 [2]第四军医大学唐都医院药剂科,陕西西安710038

出　　处：《第四军医大学学报》2005年第15期1352-1355,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30271556);全军医药卫生科研基金(02M008)

摘　　要：目的:研究互补于MRSA的耐药基因mecR1mRNA的PSODNs6088对MRSA耐药性的影响.方法:利用电穿孔的方法将PSODNs6088导入MRSA内,平板克隆形成实验计数菌落数(CFU);微量法测定细菌的生长曲线;RTPCR法检测目的基因mecR1和mecA的表达变化.结果:PSODNs6088能明显抑制MRSA的生长,PSODNs6088各剂量组MRSA的CFU与空白对照组比较明显减少(P<0.05),且具有剂量依赖性,而随机对照组的菌落数却无明显减少;PSODNs6088能显著抑制MRSAmecR1和mecA的表达.结论:反义硫代寡核苷酸能特异的与mecR1mRNA结合并阻断其表达,从而阻断了MRSA耐药基因表达信号通路,减少MRSA的菌落数,抑制其生长,提示反义寡核苷酸能有效的逆转MRSA对β内酰胺类抗生素的耐药性.利用反义技术阻断细菌的耐药信号通路可能成为解决细菌耐药问题的一种新策略.AIM： To investigate the effect of an antisense phosphothioate oligodeoxynucleotide （PS-ODNs） targeting mecRI mRNA in methicillin-resistant Staphylococcus aureus（MRSA）. METHODS： Different concentrations of PSODNs0088 were introduced into competent MRSA by electroporation. After electroporation, the total colony forming unit （CFU） per sample was counted. The cell growth curvewas drawn by measuring A600 nm at different time points with microplate reader. RT-PCR was used to observe the change of expression of mecRI and mecA in MRSA.RESULTS： Significant growth inhibition of cells treated with PS-ODNs6088 was observed as compared with those incells in control and PS-ODNs1810 treated bacteria. The number of MRSA colonies significantly decreased in all PSODNs6088 treated groups in a concentration dependent manner （ P 〈 0.05 ）, while CFU of MRSA was not influenced in mismatched PS-ODNs1810 treated group （P 〉0.05）. The expression of both mecRI and mecA was selectively inhibited. CONCLUSION： PS-ODNs targeting mecRI mRNA selectively inhibits the expression of mecRI and mecA, blocks the antibiotics resistant signal pathway and decreases the CFU of MRSA. The susceptibility of MR-SA to β-lactam antibiotics is significantly restored. The results indicate that the blockade of antibiotics resistant signal pathway via antisense approach cab be a viable strategy.

关 键 词：反义硫代寡聚脱氧核苷酸 耐甲氧西林金黄色葡萄球菌 耐药性 基因表达 

分 类 号：R378.11[医药卫生—病原生物学]