铜绿假单胞菌表面脂多糖适配体的筛选及其抑制巨噬细胞极化的研究  

Screening of aptamer binding to lipopolysaccharide of Pseudomonas aeruginosa and analysis of its inhibitory effect on macrophage polarization

作　　者：孙颖[1] 罗微[1] 张晓方 贾子超[1] 胡志东[1] 王倩[2] Sun Ying;Luo Wei;Zhang Xiaofang;Jia Zichao;Hu Zhidong;Wang Qian(Department of Clinical Laboratory, General Hospital of Tianjin Medical University, Tianjin 300052, Chin;Key Laboratory of TCM for Infectious Diseases Prevention and Control of State Administration of Traditional Chinese Medicine of the People＇s Republic of China, Department of Pathology, Tianjin Haihe Hospital, Tianjin 300050, Chin)

机构地区：[1]天津医科大学总医院检验科,300052 [2]天津市海河医院病理科国家中医药管理局中医药防治传染病重点研究室,300050

出　　处：《中华微生物学和免疫学杂志》2018年第5期354-360,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（81701968,81402391）;天津医科大学总医院青年孵育基金（ZYYFY2016024）

摘　　要：目的筛选与铜绿假单胞菌（Pseudomonas aeruginosa，PA）表面脂多糖（LPS）特异性结合的适配体，探讨其抑制巨噬细胞极化的效果。方法提取PA的LPS成分，通过指数富集的配基系统进化技术（SELEX）筛选与其特异性结合的高亲和力DNA适配体，利用酶联寡聚核苷酸吸附试验（ELONA）、定量PCR（Q-PCR）等方法探讨其对巨噬细胞极化的影响。结果本研究筛选出能与PA-LPS特异性结合的适配体PL-6，并发现其能够阻断PA-LPS与相应受体TLR4的结合，抑制巨噬细胞M1型过度极化，维持巨噬细胞稳态。结论本研究筛选出可与PA-LPS特异性结合的高亲和力适配体，为PA可能引起脓毒症的预防和治疗提供了新的策略。ObjectiveTo screen high-affinity aptamers binding to the surface lipopolysaccharide （LPS） of Pseudomonas aeruginosa （PA）, and to analyze their inhibitory effects on macrophage polarization.MethodsLPS of PA was ectracted and purified. High-affinity aptamers binding to the LPS of PA were screened by systematic evolution of ligands by exponential enrichment （SELEX）. Enzyme-linked oligonucleotide assay （ELONA） and quantitative PCR （Q-PCR） were performed to investigate their influences on macrophage polarization.ResultsIn this study, an aptamer PL-6 that could specifically bind to PA-LPS was screened successfully and found to be able to block the binding of PA-LPS to the corresponding receptor TLR4, inhibit macrophage M1 polarization and maintain macrophage homeostasis.ConclusionThis study found a high-affinity aptamer binding to the LPS of PA, which might provide a new strategy for the prevention and treatment of sepsis caused by PA.

关 键 词：铜绿假单胞菌 脂多糖 适配体 巨噬细胞 

分 类 号：R446.5[医药卫生—诊断学]