热毒宁注射液对多重耐药肺炎克雷伯菌的抗菌作用  

Antibacterial effects of Reduning Injection for multiple drug resistant Klebsiella pneumoniae

作　　者：赵娅[1,2,3] 吴娅丽 秦玺 陈小菲 王晴[2] 张明亮 王晓艳 李伟霞 唐进法[1,2,3] ZHAO Ya;WU Yali;QIN Xi;CHEN Xiaofei;WANG Qing;ZHANG Mingliang;WANG Xiaoyan;LI Weixia;TANG Jinfa(Department of Pharmacy,The First Affiliated Hospital of Henan University of Chinese Medicine,Henan Zhengzhou 450000,China;School of Pharmacy,Henan University of Chinese Medicine,Henan Zhengzhou 450046,China;Henan Province Engineering Research Center for Clinical Application,Evaluation&Transformation of Traditional Chinese Medicine,Henan Province Engineering Research Center of Safety Evaluation&Risk Management of Traditional Chinese Medicine,Henan Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院药学部,河南郑州450000 [2]河南中医药大学药学院,河南郑州450046 [3]河南省中药临床应用、评价与转化工程研究中心,河南省中药安全评价与风险防控工程研究中心,河南郑州450000

出　　处：《中国医院药学杂志》2024年第19期2243-2249,共7页Chinese Journal of Hospital Pharmacy

基　　金：河南中医药大学第一附属医院博士科研启动基金项目(编号:2021BSJJ008);河南省科技研发计划联合基金项目(编号:232301420083);河南省高等学校重点科研项目(编号:23A360008,NO.24A360020);河南省高校科技创新团队(编号:NO.23IRTSTHN026)。

摘　　要：目的:探讨热毒宁注射液(Reduning Injection,RDN)干预多重耐药肺炎克雷伯菌(multiple drug resistant Klebsiella pneumoniae,MDR-KP)的有效性。方法:收集临床分离的患者来源的KP,微量肉汤稀释法测临床常用KP防线药物美罗培南(meropenem,MEM)、替加环素(tigecycline,TIG)、多粘菌素B(colistin B,COL)的最小抑菌浓度(minimal inhibitory concentration,MIC),筛选MDR-KP,测定RDN的MIC;24 h时间-生长曲线、半定量结晶紫法分别检测RDN对KP具体生长情况的影响及对生物膜的形成和破坏作用;建立MDR-KP小鼠脓毒症模型,采用RDN进行治疗干预,检测给药后血常规、肺组织和脾组织脏器载菌量的变化,HE染色分析肺组织和脾组织的病理改变,ELISA测血清炎症因子IL-6、TNF-α、IL-1β含量。结果:MEM、TIG、COL对所选KP的MIC分别为32、64、64μg·mL–1,提示该菌株为MDR-KP。RDN对该菌株的MIC为1.3 g·mL–1。RDN对MDR-KP生长及其生物膜的形成有显著抑制作用,但对已形成的生物膜的破坏作用不明显。RDN能够调节脓毒症小鼠的白细胞总数、中性粒细胞数和单核细胞数;减少肺、脾组织脏器载菌量;修复MDR-KP造成的肺、脾组织损伤;降低血清炎症因子IL-6、TNF-α的表达。结论:RDN可通过抑制MDR-KP生长和生物膜的形成,降低脓毒症小鼠体内炎症水平,在体内外对MDR-KP均有抑制作用,既定位了RDN的临床应用范围,又丰富了RDN清热解毒的科学内涵,该研究策略也可为其他临床抗MDR-KP治疗研究提供借鉴。OBJECTIVE To explore the effectiveness of Reduning Injection(RDN)for multiple drug resistant Klebsiella pneumoniae(MDR-KP).METHODS K.pneumonia(KP)was isolated from patients.Minimal inhibitory concentrations(MICs)of meropenem(MEM),tigecycline(TIG)and colistin B(COL)were measured by broth microdilution.MDR-KP was screened and MIC of RDN determined.And 24 h time-growth curve and semi-quantitative crystal violet method were utilized for detecting the effects of RDN on specific growth of KP and formation and destruction of biofilms.MDR-KP murine septic model was established and RDN selected for therapeutic intervention.The changes of blood routine and lung/spleen organ bacterial load after dosing were detected.HE stain was employed for analyzing the pathological changes of lung and spleen tissues;The serum inflammatory factors interleukin-6(IL-6),interleukin-1 beta(IL-1β)and tumor necrosis factor-alpha(TNF-α)were detected by ELISA.RESULTS MIC of MEM,TIG,and COL for the selected KP were 32,64 and 64μg·mL–1,the strain was MDRKP.MIC of RDN was 1.3 g·mL–1.RDN had a significant inhibitory effect on the growth of MDR-KP and its biofilm formation.However,its destructive effect on formed biofilm was insignificant.RDN significantly suppressed the formation of biofilms.It could adjust the total number of leukocyte,neutrophil and monocyte in septic mice induced by MDR-KP,lower the number of bacteria in lung/spleen,repair lung/spleen tissue damage caused by MDR-KP and down-regulate the expressions of such inflammatory factors as IL-6 and TNF-α.CONCLUSION RDN may arrest the growth of MDR-KP,stunt the formation of biofilm and adjust the levels of inflammatory factors in septic mice.It exerts potent antibacterial effects on MDR-KP in vitro and in vivo,defining the clinical application scope of RDN and enriching its scientific connotation of heat-clearing and detoxification.This research strategy also provides references for other clinical trials of anti-MDR-KP treatment.

关 键 词：热毒宁注射液 多重耐药肺炎克雷伯菌 抗菌作用 生物膜 脓毒症 

分 类 号：R285.6[医药卫生—中药学] R963[医药卫生—中医学]