肝脏灌洗液中单个核细胞的抗菌作用研究  

Study of antibacterial effect of mononuclear cells in liver lavage solution

作　　者：曹薇薇[1,2] 李霄 徐纪茹[1] Cao Weiwei Li Xiao Xu Jiru(Department of Immunology and Pathogenic Biology, Medical College of Xi＇an Jiaotong University, Xi ＇an 710061, China)

机构地区：[1]西安交通大学医学院免疫学与病原生物学系,710061 [2]陕西省安康职业技术学院基础医学系 [3]第四军医大学第一附属医院肝胆外科

出　　处：《器官移植》2017年第1期27-33,共7页Organ Transplantation

基　　金：国家自然科学基金青年项目(81300361)

摘　　要：目的明确肝脏灌洗液中的单个核细胞(MC)的抗菌作用。方法体外实验:收集SD大鼠肝脏灌洗液中的MC细胞并分为两组,一组在培养液中添加白细胞介素(IL)-15,另一组不添加IL-15。将两组细胞分别与铜绿假单胞菌(P.aeruginosa)共培养4 h后收集上清并裂解细胞。以LB平板培养检测裂解液中细菌负荷,酶联免疫吸附试验(ELISA)检测培养上清中干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α含量。体内实验:将40只SD大鼠自气管注入P.aeruginosa菌液(1×109 CFU/m L)并随机分为4组,每组10只:对照组,生理盐水灌胃;免疫抑制组,他克莫司(FK506)灌胃;MC细胞组,应用FK506后自尾静脉注射MC细胞1.0×108;IL-15预处理MC细胞组,应用FK506后自尾静脉注射IL-15预处理的MC细胞1.0×108。收集各组大鼠肺组织及肺泡灌洗液,以LB平板培养检测肺组织及肺泡灌洗液中的细菌负荷,ELISA、Western blot检测肺泡灌洗液、肺组织中IFN-γ和TNF-α含量。结果与单纯MC细胞比较,IL-15预处理的MC细胞体外抗菌能力显著提高,CFU仅为前者35%,且合成、释放IFN-γ、TNF-α的能力更强。与对照组大鼠比较,免疫抑制组大鼠肺脏内免疫细胞数量减少,肺组织和肺泡灌洗液中的细菌负荷量明显升高,而IFN-γ和TNF-α含量呈下降趋势。输入IL-15预处理MC细胞后可显著提高肺组织内免疫细胞数量,减轻细菌负荷量并增加IFN-γ和TNF-α分泌。结论肝脏灌洗液中的MC细胞具有良好的抗菌活性,在免疫抑制条件下可有效提升机体对机会性致病菌的免疫防御能力。Objective To evaluate the antibacterial effect of mononuclear cells （ MCs ） in the liver lavage solution. Methods For in vitro experiment, MCs were collected from the liver lavage solution of SD rats and divided into the supplement of interleukin （ IL ） -15 and non-supplement groups. The MCs were co-cultured with Pseudomonas aeruginosa （ P aeruginosa ） for 4 h and then the supernatant was collected and MCs were lysed. The bacterial load in the lysate was detected after LB plate culture. The levels of interferon （ IFN ） -7 and tumor necrosis factor （ TNF ） -α in the supernatant were measured by enzyme-linked immune absorbent assay （ ELISA ） . For in vivo experiment, 40 SD rats were administered via tracheal injection ofP aeruginosa solution at a dose of 1 × 10^9 CFU/mL and randomly divided into four groups （ n=10 ） . In the control group, physiological saline was given via gavage. In the immunosuppression group, tacrolimus （ FK506 ） was delivered via gavage. In the MC group, MCs at a dose of 1.0 × 10^8 was given via intravenous injection after use of FK506. In the IL-15 pretreated-MC group, IL-15 pretreated-MCs at a dose of 1.0 × 10^8 were administered via intravenous injection after application of FK506. The lavage solution of pulmonary alveolus and the rat lung tissue were collected. The bacterial load was detected after LB plate culture. The expression of IFN-7 and TNF-a in the pulmonary alveolus and lung tissue were measured by ELISA and Western blot. Results Compared with MCs alone, IL-15 pretreated-MCs exhibited significantly higher antibacterial capability in vitro. The CFU was 35% of untreated MCs. The synthesis and release capabilities of IFN-T and TNF-× were significantly enhanced. Compared with the control group, the quantity of immune cells in the lung tissue was decreased and the bacterial load in the lung tissue and the lavage solution of pulmonary alveolus was significantly increased, whereas the expression levels of IFN-7 and TNF-× tended to decline in the imm

关 键 词：铜绿假单胞菌 肝脏 灌洗液 单个核细胞 抗感染 

分 类 号：R617[医药卫生—外科学] R378.991[医药卫生—临床医学]