黄连等五种中药颗粒剂对鲍曼不动杆菌生物膜耐药性及相关基因的作用分析

Analysis of the Effect of Five Kinds of Traditional Chinese Medicine Granules Including Huanglian on the Biofilm Resistance of Acinetobacter Baumannii and Related Genes

目的:评估碳青霉烯类耐药鲍曼不动杆菌(CRAB)的耐药基因受黄连等五种中药颗粒的影响。方法:对广东省第二中医院收集的碳青霉烯类耐药鲍曼不动杆菌进行试验,结晶紫染色法挑选生物膜阳性菌株2株;在体外建立鲍曼不动杆菌生物膜模型,通过肉汤倍比稀释法测定黄连等5种中药颗粒剂对产膜原菌最小抑膜浓度;留取抑膜亚MIC浓度实验菌液进行常见耐药基因检测及药敏实验,对比用药前后的药敏结果及常见18对耐药基因检测结果的改变。结果:耐药基因检测主要改变为,药物作用前原菌液检出floR基因,用药后检出arm基因、sul基因及floR基因,药物作用前后菌液抗菌药物药敏结果复方新诺明和替加环素对实验菌株MIC值有不同程度改变。结论:黄连等五种中药颗粒剂能够不同程度抑制碳青霉烯类耐药鲍曼不动杆菌生物膜形成,通过测定用药前后抗菌药物敏感性实验以及常见耐药基因,发现中药能够对细菌内叶酸合成途径以及细菌蛋白质形成产生影响;综合用药前后药敏结果及基因检测结果,药敏与碳青霉烯类耐药鲍曼不动杆菌是否形成生物膜有差异、中药颗粒剂种类无显著差异,可能中药抑膜、抑菌作用机制与抗生素作用机制有差异。

Objective:To evaluate the effect of drug resistance genes of carbapenem-resistant Acinetobacter baumannii(CRAB)on five kinds of traditional Chinese medicine granules including Huanglian.Methods:The carbapenem-resistant Acinetobacter baumannii collected in the Second Hospital of Traditional Chinese Medicine of Guangdong Province was tested,and 2 biofilm-positive strains were selected by crystal violet staining;the biofilm model of Acinetobacter baumannii was established in vitro,The minimum inhibitory concentration of 5 kinds of traditional Chinese medicine granules,including Coptis chinensis,on film-producing bacteria was determined by the broth multi-dilution method;Drug susceptibility results and changes in the detection results of common 18 pairs of drug resistance genes.Results:The main changes in the detection of drug resistance genes were that the floR gene was detected in the original bacterial solution before the drug action,the arm gene,sul gene and floR gene were detected after the drug treatment,and the antibacterial drug susceptibility results of the bacterial solution before and after the drug action.The MIC values of the experimental strains have different degrees of changes.Conclusion:Five kinds of traditional Chinese medicine granules such as Huanglian can inhibit the biofilm formation of carbapenem-resistant Acinetobacter baumannii to different degrees.Folic acid synthesis pathway and bacterial protein formation are affected;the drug susceptibility results and genetic testing results before and after the drug are combined,and there is a difference between the drug susceptibility and carbapenem-resistant Acinetobacter baumannii whether biofilm is formed,and there is no significant difference in the types of traditional Chinese medicine granules,it is possible that the mechanism of Chinese medicine’s anti-membrane and bacteriostatic action is different from that of antibiotics.