痰热清注射液联合阿米卡星对铜绿假单胞菌耐药菌株体外抗菌作用研究
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摘要
为了研究痰热清注射液联合阿米卡星对铜绿假单胞菌耐药菌株的体外抗菌作用,试验采用微量稀释法分别测定痰热清注射液、阿米卡星以及两药联合对铜绿假单胞菌耐药菌株1号、耐药菌株2号的最低抑菌浓度(MIC);采用棋盘稀释法检测两药联合的体外抑菌作用,并计算部分抑菌浓度(FIC)指数,根据FIC指数判定药物联合的抗菌作用;建立体外生物膜模型,用银染法分别测定阿米卡星、痰热清注射液及两药联合对其成膜能力的影响。结果表明:对于耐药菌株1号,阿米卡星的MIC为6.25μg/mL,痰热清注射液的MIC为62.5μL/mL,联合用药时阿米卡星的MIC为1.56μg/mL,痰热清注射液的MIC为1.95μL/mL;对于耐药菌株2号,阿米卡星的MIC为12.5μg/mL,痰热清注射液的MIC为500μL/mL,联合用药时阿米卡星的MIC为0.39μg/mL,痰热清注射液的MIC为125μL/mL;对于两个耐药菌株,两药联合时的FIC指数均为0.28。两个耐药菌株的成膜能力强,高浓度的阿米卡星和痰热清注射液单独使用时均对其成膜能力有抑制作用,而低浓度阿米卡星反而有促进生物膜生长的作用(阿米卡星浓度≤3.125μg/mL促进耐药菌株1号生物膜生长,阿米卡星浓度≤6.25μg/mL促进耐药菌株2号生物膜生长),痰热清注射液与阿米卡星联合用药后对两个耐药菌株生物膜生长的抑制作用增强。说明痰热清注射液与阿米卡星联合对耐药菌株1号和耐药菌株2号的抑制具有协同作用。
To study the antibacterial effect of Tanreqing injection combined with amikacin on the drug-resistant strains of Pseudomonas aeruginosa,the minimum inhibitory concentration(MIC) was determined by brouth dilution method.The fractional inhibitory concentration(FIC) index was detected to study the joint antibacterial effect of combination of drugs.In vitro, biofilm model was established to determine the effect of amikacin, Tanreqing injection and the combination of two drugs on the film-forming ability.The result showed that,as todrug-resistant strain 1, the MIC of amikacin was 6.25 μg/mL, the MIC of Tanreqing injection was 62.5 μL/mL; when they are mixed, the MIC of amikacin was 1.56 μg/mL, and the MIC of Tanreqing injection was 1.95 μL/mL. As to drug-resistant strain 2, the MIC of amikacin was 12.5 μg/mL, the MIC of Tanreqing injection was 500 μL/mL;when they are mixed, the MIC of of amikacin was 0.39 μg/mL, the MIC of Tanreqing injection was 125 μL/mL.The FIC index of combined drugs on strain 1 or strain 2 was 0.28.These two drug-resistant strains had the high ability to form biofilm. Higher concentration of amikacin or Tanreqing injection has inhibited the biofilm growth. However, the low concentration of amikacin can promote the growth of biofilm, with the concentration below 3.125 μg/mL on strain 1,and with the concentration below 6.25 μg/mL on strain 2. The combination of Tanreqing injection and amikacin has inhibited the biofilm growth.It indicated that combination of Tanreqing injection and amikacin had synergism on resistant strain 1 or drug-resistant strain 2.
To study the antibacterial effect of Tanreqing injection combined with amikacin on the drug-resistant strains of Pseudomonas aeruginosa,the minimum inhibitory concentration(MIC) was determined by brouth dilution method.The fractional inhibitory concentration(FIC) index was detected to study the joint antibacterial effect of combination of drugs.In vitro, biofilm model was established to determine the effect of amikacin, Tanreqing injection and the combination of two drugs on the film-forming ability.The result showed that,as todrug-resistant strain 1, the MIC of amikacin was 6.25 μg/mL, the MIC of Tanreqing injection was 62.5 μL/mL; when they are mixed, the MIC of amikacin was 1.56 μg/mL, and the MIC of Tanreqing injection was 1.95 μL/mL. As to drug-resistant strain 2, the MIC of amikacin was 12.5 μg/mL, the MIC of Tanreqing injection was 500 μL/mL;when they are mixed, the MIC of of amikacin was 0.39 μg/mL, the MIC of Tanreqing injection was 125 μL/mL.The FIC index of combined drugs on strain 1 or strain 2 was 0.28.These two drug-resistant strains had the high ability to form biofilm. Higher concentration of amikacin or Tanreqing injection has inhibited the biofilm growth. However, the low concentration of amikacin can promote the growth of biofilm, with the concentration below 3.125 μg/mL on strain 1,and with the concentration below 6.25 μg/mL on strain 2. The combination of Tanreqing injection and amikacin has inhibited the biofilm growth.It indicated that combination of Tanreqing injection and amikacin had synergism on resistant strain 1 or drug-resistant strain 2.
引文
[1] 杨启文,王辉,徐英春,等.环丙沙星或阿米卡星联合β-内酰胺类抗生素对多重耐药铜绿假单胞菌的体外联合抑菌效应研究[J].中国实用内科杂志,2006(9):685-687.
[2] 王越,张冬,王慧敏,等.痰热清注射液对铜绿假单胞菌生物被膜形成的影响[J].中华医院感染学杂志,2016,26(21):4841-4843,4858.
[3] 晋兴,武爱荣,黄波,等.用纸片扩散法和微量稀释法进行药物敏感性测定的比较[J].国际检验医学杂志,2014,35(3):322-324.
[4] 宋延君,张宏,李水秀,等.以棋盘法测定并评价汉防己甲素与酮康唑联合对白念珠菌的体外抗真菌活性[J].中国人兽共患病学报,2012,28(9):890-893.
[5] 邵圣文,张志智,顾红光,等.RNA干扰铜绿假单胞菌群体感应系统对生物膜影响的研究[J].中国抗生素杂志,2009,34(3):184-188.
[6] 李晓霞,郭阳,张瑞琴,等.铜绿假单胞菌耐药率与生物膜形成能力之间的相关性分析[J].中华医院感染学杂志,2014,24(12):2868-2870,2879.
[7] 赵宁波. 痰热清注射液对细菌生物膜影响的研究[N]. 中国中医药报,2014-04-18(7).
[8] 王越,张冬,王慧敏,等.痰热清注射液对铜绿假单胞菌生物被膜形成的影响[J].中华医院感染学杂志,2016,26(21):4841-4843,4858.
[9] 李俊娟,王强,孙开宇,等.铜绿假单胞菌生物膜与亚抑菌浓度抗菌药物的相关研究[J].中华医院感染学杂志,2012,22(24):5437-5440.
[10] 孔晋亮,刘晓岚,陈一强,等.银染法观察铜绿假单胞菌生物被膜变化初探[J].济宁医学院学报,2007,30(1):27-29.
[11] 李苏利,华川.铜绿假单胞菌多重耐药及泛耐药研究进展[J].山西医药杂志,2013,42(8):891-892.
[12] 汪长中.中药抗细菌生物膜研究进展[J].中国中药杂志,2010,35(4):521-524.
[13] 韩燕鸿,张荷.痰热清注射液治疗小儿上呼吸道感染的系统评价[J].中国实验方剂学杂志,2016,22(12):215-219.
[14] 朱丽莎,艾彪,杜昆,等.肠杆菌科细菌对头孢哌酮/舒巴坦的耐药性分析[J].中华医院感染学杂志,2012,22(4):809-810.
[2] 王越,张冬,王慧敏,等.痰热清注射液对铜绿假单胞菌生物被膜形成的影响[J].中华医院感染学杂志,2016,26(21):4841-4843,4858.
[3] 晋兴,武爱荣,黄波,等.用纸片扩散法和微量稀释法进行药物敏感性测定的比较[J].国际检验医学杂志,2014,35(3):322-324.
[4] 宋延君,张宏,李水秀,等.以棋盘法测定并评价汉防己甲素与酮康唑联合对白念珠菌的体外抗真菌活性[J].中国人兽共患病学报,2012,28(9):890-893.
[5] 邵圣文,张志智,顾红光,等.RNA干扰铜绿假单胞菌群体感应系统对生物膜影响的研究[J].中国抗生素杂志,2009,34(3):184-188.
[6] 李晓霞,郭阳,张瑞琴,等.铜绿假单胞菌耐药率与生物膜形成能力之间的相关性分析[J].中华医院感染学杂志,2014,24(12):2868-2870,2879.
[7] 赵宁波. 痰热清注射液对细菌生物膜影响的研究[N]. 中国中医药报,2014-04-18(7).
[8] 王越,张冬,王慧敏,等.痰热清注射液对铜绿假单胞菌生物被膜形成的影响[J].中华医院感染学杂志,2016,26(21):4841-4843,4858.
[9] 李俊娟,王强,孙开宇,等.铜绿假单胞菌生物膜与亚抑菌浓度抗菌药物的相关研究[J].中华医院感染学杂志,2012,22(24):5437-5440.
[10] 孔晋亮,刘晓岚,陈一强,等.银染法观察铜绿假单胞菌生物被膜变化初探[J].济宁医学院学报,2007,30(1):27-29.
[11] 李苏利,华川.铜绿假单胞菌多重耐药及泛耐药研究进展[J].山西医药杂志,2013,42(8):891-892.
[12] 汪长中.中药抗细菌生物膜研究进展[J].中国中药杂志,2010,35(4):521-524.
[13] 韩燕鸿,张荷.痰热清注射液治疗小儿上呼吸道感染的系统评价[J].中国实验方剂学杂志,2016,22(12):215-219.
[14] 朱丽莎,艾彪,杜昆,等.肠杆菌科细菌对头孢哌酮/舒巴坦的耐药性分析[J].中华医院感染学杂志,2012,22(4):809-810.