绿原酸、c-di-GMP、人杀菌肽LL-37对铜绿假单胞菌生物被膜耐药性的影响  

Effect of chlorogenic acid,c-di-GMP and human bactericidal peptide LL-37 on drug resistance of Pseudomonas aeruginosa biofilm

作　　者：张艳 赵淑珍 何静 张婷 ZHANG Yan;ZHAO Shu-zhen;HE Jing;ZHANG Ting(Department of Clinical Laboratory, Urumqi Hospital of Traditional Chinese Medicine, Xinjiang Uygur Autonomous Region, Urumqi 830000, China)

机构地区：[1]新疆维吾尔自治区乌鲁木齐市中医医院检验科,新疆乌鲁木齐830000

出　　处：《河北医科大学学报》2022年第6期721-726,共6页Journal of Hebei Medical University

基　　金：乌鲁木齐市卫生健康委科技计划项目(202039)。

摘　　要：目的 探究铜绿假单胞菌(pseudomonas aeruginosa,PA)的生物被膜(Biofilm,BF)形成与耐药性中绿原酸(Chlorogenic acid)、c-di-GMP、人杀菌肽LL-37的作用。方法 选取铜绿假单胞菌野生型株(pseudomonas aeruginosa strain,PAO1)展开研究,采用肉汤稀释法测定绿原酸、c-di-GMP、人杀菌肽LL-37对PA最低抑菌浓度(minimum inhibitory concentration,MIC),采用结晶紫定量法检测PA的BF形成中绿原酸、c-di-GMP、人杀菌肽LL-37的清除作用,采用PA运动试验检测绿原酸、c-di-GMP、人杀菌肽LL-37对PA运动能力的影响,荧光显微镜下观察绿原酸、c-di-GMP、人杀菌肽LL-37对PA的BF结构及存活情况的影响。结果 人杀菌肽LL-37MIC值为64 mg/L、绿原酸MIC值为256 mg/L、c-di-GMPMIC值为32 mg/L,相比于c-di-GMP组,人杀菌肽LL-37组、绿原酸组、空白组的OD590nm值明显降低,且人杀菌肽LL-37组、绿原酸组明显降低(P<0.05)。相比阴性对照组,人杀菌肽LL-37组、绿原酸组、c-di-GMP组的PA运动试验直径均明显降低,且相比于c-di-GMP组,人杀菌肽LL-37组、绿原酸组明显降低(P<0.05)。空白组、c-di-GMP组的PA存活情况优于人杀菌肽LL-37组、绿原酸组,空白组、c-di-GMP组主要为绿色荧光,人杀菌肽LL-37组、绿原酸组主要为红色荧光、橙色荧光。相比c-di-GMP组,人杀菌肽LL-37组、阴性对照组、绿原酸组的耐药率明显降低,且相比于阴性对照组,人杀菌肽LL-37组、绿原酸组明显降低(P<0.05)。结论 绿原酸、人杀菌肽LL-37对PA的BF均可以发挥一定的清除作用,c-di-GMP可通过抑制细胞运动能力对PA的BF形成起到一定的调控作用,绿原酸、人杀菌肽LL-37以及c-di-GMP均可作为后续防治PA的BF的新方向。Objective To investigate the role of chlorogenic acid,c-di-GMP and human bactericidal peptide LL-37 in biofilm formation(BF)of Pseudomonas aeruginosa(PA)and drug resistance.Methods Wild-type pseudomonas aeruginosa strain(PAO1)was selected to carry out the study,and the minimum inhibitory concentration(MIC)of chlorogenic acid,c-di-GMP and human germicide LL-37 on PA was determined by broth dilution method.Crystal violet quantitative method was used to detect the scavenging effects of chlorogenic acid,c-di-GMP and human germicidal peptide LL-37 in PA-BF formation,and PA motion test was used to detect the effects of chlorogenic acid,c-di-GMP and human germicidal peptide LL-37 on PA motion ability.The effects of chlorogenic acid,c-di-GMP and human bactericidal peptide LL-37 on the structure and survival of PA-BF were observed under fluorescence microscope.Results The value of MIC of human germicidal peptide LL-37,chlorogenic acid and c-di-GMP was 64 mg/L,256 mg/L,and 32 mg/L,respectively.Compared with the c-divGMP group,the value of OD590nm human germicidal peptide LL-37 group,chlorogenic acid group and blank group significantly decreased,which was significantly lower in human bactericidal peptide LL-37 group and chlorogenic acid group(P<0.05).Compared with the negative control group,the PA diameter in exercise testing of the human germicidal peptide LL-37 group,the chlorogenic acid group and the c-di-GMP group significantly decreased,which was significantly lower in human germicidal peptide LL-37 group and the chlorogenic acid group,as compared with the c-di-GMP group(P<0.05).The survival of PA in blank group and c-divGMP group was better than that in human germicidal peptide LL-37 group and chlorogenic acid group.The fluorescence in blank group and c-di-GMP group was mainly green,while the fluorescence in human germicidal peptide LLv37 group and chlorogenic acid group was mainly red and orange.Compared with c-di-GMP group,the drug resistance rate of human germicidal peptide LL-37 group,negative control group and ch

关 键 词：铜绿假单胞菌 生物被膜 耐药性 

分 类 号：R378.991[医药卫生—病原生物学]