TLR2配基Pam3CSK4对小鼠中性粒细胞吞噬耐甲氧西林金黄色葡萄球菌功能的影响  

Effects of TLR2 Ligand Pam3CSK4 on the Function of Mouse Neutrophil Phagocytic Methicillin-resistant Staphylococcus aureus

作　　者：陆姗姗[1] 陈益国[2] 张咏 邓林强[2] 陈会[2] 闵卫平 LU Shan-shan;CHEN Yi-guo;ZHANG Yong;DENG Lin-qiang;CHEN Hui;MIN Wei-ping(Department of Pharmacy, Nanchang University Medical College and Institute of lmmunotherapy, Nanchang University, Nanchang 330006, Chin)

机构地区：[1]南昌大学医学院药学系,南昌大学免疫学与生物治疗研究所,江西南昌330006 [2]江西省人民医院,江西南昌330008 [3]江西省医学科学研究院,江西南昌330006

出　　处：《现代预防医学》2018年第8期1453-1457,1482,共6页Modern Preventive Medicine

基　　金：国家自然科学基金(81360454); 南昌大学研究生创新专项资金(cx2016303)

摘　　要：目的探讨Toll样受体2(TLR2)配基Pam3CSK4对小鼠骨髓来源中性粒细胞吞菌功能的影响。方法取小鼠骨髓中性粒细胞,给予Pam3CSK4或LTA刺激,采用流式细胞术检测中性粒细胞对热灭活的耐甲氧西林金黄色葡萄球菌(MRSA),耐碳青霉烯类的鲍曼不动杆菌(CR-AB),耐碳青霉烯类的肺炎克雷伯菌(CR-KP)的吞噬作用;磁珠分选中性粒细胞,给予Pam3CSK4刺激24 h、48 h后,q-PCR检测中性粒细胞IL-1β,IL-6,IL10,TNF-α,FcγRⅢ,i NOS,COX-2等基因表达水平的改变;Pam3CSK4刺激中性粒细胞不同时间后再给予热灭活的MRSA刺激,收集培养上清液,ELISA检测IL-1,IL-6,TNF-α细胞因子的表达水平。结果 Pam3CSK4刺激组的中性粒细胞对热灭活MRSA的吞噬率为41.1%,明显高于PBS对照组(18.4%),但Pam3CSK4刺激组对CR-AB和CR-KP的吞噬率与对照组相比没有明显改变。Pam3CSK4刺激中性粒细胞12 h,24 h后IL-6和TNF-α的表达与PBS对照组相比明显增加,差异有统计学意义(P<0.05),此外,Pam3CSK4刺激组增强中性粒细胞IL-1β、IL-6、IL10、TNF-α、FcγRⅢ、i NOS、COX-2的m RNA的表达(P<0.05)。结论 Pam3CSK4可以增强中性粒细胞对MRSA的吞噬作用。Objective To investigate the effect of Toll like receptor 2（TLR2） ligand Pam3 CSK4 on the phagocytosis of murine bone marrow derived neutrophils.Methods The mice bone marrow neutrophils were treated with Pam3 CSK4 or LTA,and flow cytometry was used to detect the phagocytosis of neutrophils against heat-killed methicillin-resistant Staphylococcus aureus（MRSA）,carbapenem resistant Acinetobacter Bauman（CR-AB） and carbapenem resistant Klebsiella pneumoniae（CRKP）.N eutrophils were purified with magnetic beads and stimulated with Pam3 CSK4 for 24 h or 48 h,and then the gene expressions of IL-1,IL-6,IL10,TNF-α,FcγRⅢ,i NOS,COX were detected by q-PCR; Pam3 CSK4 pretreated neutrophils were given MRSA stimulation at different time,the supernatant was collected and the protein levels of IL-1,IL-6,TNF-αwere detected by ELISA.Results The phagocytosis rate of neutrophils to heat-killed MRSA in Pam3 CSK4 stimulation group was 41.1%,which was significantly higher than that in PBS control group（18.4%）,but the phagocytosis rate of CR-AB and CR-KP in Pam3 CSK4 stimulation group did not change significantly compared with that in control group.Pam3 CSK4 stimulated neutrophils for 12 h and 24 h significantly increased the expression of IL-6 and TNF-α compared with that in PBS control group（P〈0.05）.In addition,Pam3 CSK4 stimulation group enhanced the expression of IL-1β,IL-6,IL10,TNF-α,FcγRⅢ,i NOS,and COX-2 at gene level.Conclusion Pam3 CSK4 pretreatment can enhance the phagocytosis of neutrophils to MRSA.

关 键 词：Pam3CSK4 MRSA 中性粒细胞 

分 类 号：R392.11[医药卫生—免疫学]