可利霉素对堪萨斯分枝杆菌的体内外抗菌活性研究  

Study on the antibacterial activity of carrimycin against Mycobacterium Kansasii in vitro and in vivo

作　　者：王梓墨 李晓东 张炜焱 付雷 陈曦 陈效友 陆宇 Wang Zimo;Li Xiaodong;Zhang Weiyan;Fu Lei;Chen Xi;Chen Xiaoyou;Lu Yu(Beijing Key Laboratory of Drug Resistance Tuberculosis Research,Beijing Chest Hospital,Capital Medical University,Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149;Beijing Ditan Hospital,Capital Medical University,Beijing 100015)

机构地区：[1]首都医科大学附属北京胸科医院药物学研究室,北京市结核病胸部肿瘤研究所,耐药结核病北京市重点实验室,北京101149 [2]首都医科大学附属北京地坛医院,北京100015

出　　处：《中国抗生素杂志》2024年第12期1374-1383,共10页Chinese Journal of Antibiotics

基　　金：北京市医院管理中心“登峰”人才培养计划(No.DFL20221402);首都卫生科研发展专项项目(No.首发2022-1-2171)。

摘　　要：目的评价新一代大环内酯类药物可利霉素(carrimycin,CAM)对堪萨斯分枝杆菌(Mycobacterium kansasii,MK)的体内外抗菌活性,旨在为治疗堪萨斯分枝杆菌引起的感染提供更多选择。方法选取MK标准株(ATCC12478)和3株临床分离株作为研究菌株,通过微孔板显色法(microplate alamar blue assay,MABA)测定CAM、螺旋霉素(spiramycin,SPM)、阿奇霉素(azithromycin,AZM)和利福平(rifampicin,RIF)的最低抑菌浓度(minimal inhibitory concentration,MIC);棋盘法测定CAM与AZM和RIF等8种临床常用治疗药物的体外作用;评估CAM、SPM、AZM在J774A.1细胞中的抗菌活性;建立BALB/c小鼠感染堪萨斯分枝杆菌的模型,并通过CFU计数评估空白对照组、CAM治疗组、AZM治疗组和SPM治疗组小鼠脏器活菌数,苏木精-伊红染色法(HE染色)评价CAM治疗组小鼠肺组织病变程度。结果CAM、SPM、AZM和RIF在MK标准株(ATCC12478)和3株临床分离株中测定的MIC值分别为4~8 mg/L、>64 mg/L、4~8 mg/L和0.125~0.25 mg/L。8种临床常用治疗药物和CAM在体外未出现拮抗作用(fractional inhibitory concentration index,FICI<4),CAM和异烟肼、利福平等药物具有部分协同作用。在巨噬细胞J774A.1感染MK实验中,和对照组相比2 mg/L CAM、SPM和AZM处理48 h均能使J774A.1细胞内MK降低0.3~0.4 lgCFU(P<0.001)。在感染中等剂量MK的BALB/c小鼠模型中给与100 mg/kg药物进行治疗,与空白对照组相比CAM、AZM和SPM治疗2周小鼠肺组织中活菌数分别降低了0.73、0.69和0.22 lgCFU(P<0.001),治疗4周小鼠肺组织中活菌数分别降低了0.74、1.02和0.41 lgCFU(P<0.001)。在感染高剂量MK的BALB/c小鼠模型中,与空白对照组相比100 mg/kg CAM治疗4周小鼠肺组织中活菌数降低了0.98 lgCFU(P<0.001),HE染色结果显示CAM治疗显著降低了小鼠肺组织中炎症细胞数量(P=0.001)。结论CAM对MK具有较好的体内外抗菌活性,可以显著降低肺组织活菌数,减轻肺组织炎症程度,值得进一步开展临床研Objective To evaluate the activity of carrimycin(CAM)against M.kansasii(MK)in vitro and in vivo and provide more options for clinical treatment in pulmonary disease due to MK.Methods The standard strain(ATCC12478)and 3 clinical isolates of MK were used in experiments.The microplate alamar blue assay(MABA)was used to determine the minimal inhibitory concentrations(MICs)of CAM,spiramycin(SPM),azithromycin(AZM)and rifampicin(RIF).The checkerboard assay was used to evaluate the interaction between CAM and 8 clinically used drugs including AZM and RIF.The antibacterial activity of CAM,AZM and SPM in J774A.1 cells was evaluated and the model of BALB/c mice infected with MK was established.CFU counting was used to determine the bacillary load of organs in the black control group,CAM treatment group,AZM treatment group,and SPM treatment group.HE staining was used to assess the degree of lung injury.Results The MIC values of CAM,SPM,AZM and RIF in MK standar strains(ATCC12478)and three clincal isolates were 4~8 mg/L,>64 mg/L,4~8 mg/L and 0.125~0.25 mg/L,respectively.No antagonism between CAM and drugs used in this study were observed(fractional inhibitory concentration index,FICI<4).CAM was partially synergized with isoniazid and rifampicin.The concentration of 2 mg/L CAM,SPM and AZM could reduce lgCFU from 0.3 to 0.4 in the macrophage infection model compared with the control group(P<0.001).In the BALB/c mouse model infected with middle dosage MK,compared with the control group,continuous administration of 100 mg/kg CAM,AZM and SPM could reduce the bacillary load to 0.73,0.69 and 0.22 lgCFU(P<0.001)in the lungs after 2 weeks of treatment and 0.74,1.02 and 0.41 lgCFU(P<0.001)after 4 weeks of treatment,respectively.In the BALB/c mouse model infected with high-dose MK,continuous administration of 100 mg/kg CAM for 4 weeks could reduce the bacillary load 0.98 lgCFU(P<0.001)in the lungs compared with the control group.CAM treatment could significantly reduce inflammatory cells compared with the control group(P=0.001).Conclusi

关 键 词：堪萨斯分枝杆菌 非结核分枝杆菌 可利霉素 阿奇霉素 螺旋霉素 

分 类 号：R978[医药卫生—药品]