鱼腥草水提物和大环内酯类药物体外诱导肺炎支原体耐药的对比研究  

Comparison of water extract of Houttuynia cordata and macrolide drugs to induce resistance of Mycoplasma pneumoniae isolates in vitro

作　　者：凌琳 金秀萍 黄燕颖 孙爱华 Ling Lin;Jin Xiuping;Huang Yanying;Sun Aihua(Department of Pharmacy,General Hospital of Medical Community,Keqiao District Hospital of Traditional Chinese Medicine of Shaoxing,Shaoxing 312030,Zhejiang,China;Clinical Laboratory,General Hospital of Medical Community,Keqiao District Hospital of Traditional Chinese Medicine of Shaoxing,Shaoxing 312030,Zhejiang,China;Department of Pathology,Hangzhou Red Cross Hospital,Hangzhou 310003,China;Department of Basic Medical Sciences and Forensic Medicine,Hangzhou Medical College,Hangzhou 310053,China)

机构地区：[1]绍兴市柯桥区中医医院医共体总院药剂科,浙江绍兴312030 [2]绍兴市柯桥区中医院医共体总院检验科,浙江绍兴312030 [3]杭州市红十字会医院病理科,310003 [4]杭州医学院基础医学与法医学院,310053

出　　处：《国际流行病学传染病学杂志》2021年第3期206-210,共5页International Journal of Epidemiology and Infectious Disease

基　　金：国家自然科学基金(81772232);绍兴市柯桥区社发类科技计划(2020KZ70)。

摘　　要：目的了解亚抑制浓度鱼腥草水提物和红霉素、阿奇霉素、交沙霉素对肺炎支原体(MP)大环内酯类药物敏感株的耐药诱导作用,探讨鱼腥草用于治疗MP感染的可能优势。方法10株对3种大环内酯类药物均敏感的MP临床菌株体外用不同浓度梯度的鱼腥草水提物和大环内酯类药物逐级诱导传代。用微量稀释法测定药物对传代后菌株的最低抑菌浓度。PCR扩增耐药相关基因23S rRNA结构域V区和核糖体蛋白L4、L22基因并测序。结果经红霉素、阿奇霉素、交沙霉素体外诱导后,分别有8株、5株和4株成为红霉素、阿奇霉素、交沙霉素稳定耐药株;与耐药诱导前相比,药物对各临床菌株的最低抑菌浓度分别提高了2~16倍;诱导前后,所有菌株的核糖体蛋白L4、L22编码基因测序均未发现突变,8株诱导耐药株中的6株出现23S rRNA结构域V区A2063G/T突变。经鱼腥草水提物诱导传代的各临床菌株最低抑菌浓度提高均不超过2倍。结论体外多步骤亚抑菌浓度大环内酯类药物诱导作用可致MP的23S rRNA选择性突变而产生稳定性耐药。与抗菌药物相比,鱼腥草水提物诱导后的MP不易产生耐药。Objective To investigate the sub-minimum inhibitory concentration(MIC)of water extract of Houttuynia cordata and erythromycin,azithromycin,josamycin that induce resistance in macrolide-sensitive M.pneumoniae(MP),and to explore the possible advantages of Hyouttuynia cordata in the treatment of MP infection.Methods A total of 10 isolates of MP that were sensitive to all three macrolide drugs were induced and subcultured by Hyouttuynia cordata and macrolide drugs with different concentration gradients in vitro.The MICs of drugs to the subcultured strains were determined by microdilution.The 23S rRNA V region and ribosomal protein L4 and 122 genes related to drug resistance were amplified by PCR and sequenced.Results After induction of erythromycin,azithromycin and josamycin in vitro,there were 8,5 and 4 stable isolates resistant to erythromycin,azithromycin and josamycin,respectively.The MICs of MP isolates induced by macrolide drugs increased 2-16 times,respectively.The sequencing results showed that L4 and L22 encoding genes of all isolates had no mutant before and after induction.However,6 of the 8 induced resistant isolates had mutation of A2063G/T in 23S rRNA domain V region.The MICs of all clinical strains induced by water extract of Hyouttuynia cordata increased within 2 times.Conclusions The multi-step induction of macrolides with sub-MICs in vitro can lead to the selective mutation of 23S rRNA and stable drug resistance of MP.Compared with antimicrobial agents,MP induced by water extract of Hyouttuynia cordata has less acquired resistance.

关 键 词：支原体 肺炎 鱼腥草 大环内酯类药物 体外 诱导耐药 基因突变 

分 类 号：R965[医药卫生—药理学]