摘要
目的探讨双黄连影响兔组织笼感染模型中左氧氟沙星对金黄色葡萄球菌grlA、grlB、gyrA、gyrB基因突变的影响,以期为中药改善喹诺酮类药物耐药提供科学依据。方法建立金黄色葡萄球菌ATCC 29213感染兔组织笼模型,分别以生理盐水,单用左氧氟沙星(10mg·kg^(-1)·d^(-1)),单用双黄连(85mg·kg^(-1)·d^(-1)),左氧氟沙星(10mg·kg^(-1)·d^(-1))联合低、中、高剂量双黄连(42.5、85、170mg·kg^(-1)·d^(-1))干预,连续给药5d,提取干预结束后两天组织笼内恢复生长的细菌DNA,扩增,测定分析grlA、grlB、gyrA、gyrB基因序列,比较各基因序列突变情况与细菌耐药性的关系。结果单用左氧氟沙星处理后细菌grlA基因序列发生多处突变,grlB、gyrA基因序列发生了两处点突变,而gyrB未发生突变;单用双黄连处理、双黄连联合左氧氟沙星处理后细菌上述基因均未发生突变。结论左氧氟沙星引起细菌耐药性的产生可能与grlA基因中的多处点突变,grlB、gyrA的两处点突变有关,可能与gyrB基因无关。而双黄连能抑制左氧氟沙星诱导产生的金黄色葡萄球菌grlA、grlB、gyrA的突变。
OBJECTIVE To explore the effect of Shuanghuanglian on levofloxacin-induced grlA,grlB,gyrA and gyrBgenes mutations of Staphylococcus aureus in rabbit tissue cage infection model so as to provide scientific basis for improvement of quinolones resistance by traditional Chinese Medicine.METHODS The tissue cage infection models with S.aureus were established in rabbits and were respectively given normal saline injection,single levofloxacin(10mg·kg-1·d-1)and levofloxacin(10mg·kg-1·d-1)combined with low-,medium-and high-dose Shuanghuanglian(42.5,85,170mg·kg-1·d-1)for 5 consecutive days,DNA were extracted from the bacteria that grew in the tissue inside the cage after the intervention for 2 days and were amplified,the grlA,grlB,gyrA and gyrBgenes were sequenced,the mutations of the genes were compared,and the correlation between the mutations and the drug resistance was observed.RESULTS The grlAgene sequence had multiple mutations after the models were treated with single levofloxacin,the grlB and gyrA gene sequences had the mutations in two loci,while the gyrBdid not have mutations.The above genes did not have mutations after the models were treated with single Shuanghuanglian and Shuanghuanglian combined with levofloxacin.CONCLUSIONThe levofloxacin-induced drug resistance may be associated with the multiple mutations of the grlA gene and the two loci of mutations in grlBand gyrA genes but is not associated with the mutations of the gyrBgene.Shuanghuanglian may inhibit the levofloxacin-induced mutations of grlA,grlBand gyrA genes in the S.aureus.
作者
罗恒丽
代颜
陈竹
王国俊
LUO Heng-li;DAI Yan;CHEN Zhu;WANG Guo-jun(The Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000,China)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2019年第9期1281-1287,共7页
Chinese Journal of Nosocomiology
基金
四川省医学科研青年创新基金资助项目(Q14032)
关键词
兔组织笼感染模型
左氧氟沙星
双黄连
金黄色葡萄球菌
基因突变
Rabbit tissue cage infection model
Levofloxacin
Shuanghuanglian
Staphylococcus aureus
Gene mutation
作者简介
通信作者:王国俊,E-mail:wanggj7532@163.com
