梅毒螺旋体脂肽诱导THP-1细胞表达细胞因子的研究  

The research of cytokines-induced function of Treponema pallidum lipopeptides activated THP-1

作　　者：孙冉[1] 刘子莲[2] 褚小玲[1] 张海萍[3] 

机构地区：[1]首都医科大学附属北京安贞医院,北京100029 [2]北京大学第三医院 [3]首都医科大学宣武医院

出　　处：《实用皮肤病学杂志》2015年第5期336-339,共4页Journal of Practical Dermatology

基　　金：北京市自然科学基金资助项目(7102079)

摘　　要：目的通过体外建立巨噬细胞模型,研究梅毒螺旋体(TP)相对分子质量15 000、17 000、47 000膜脂蛋白诱导THP-1细胞表达细胞因子的活性,评价其在梅毒感染细胞免疫中的作用,为进一步研究梅毒螺旋体的致病机制和生物学功能提供理论基础。方法用不同剂量PMA诱导THP-1细胞,分别观察细胞的形态和检测膜表面CD14受体表达情况,确定PMA诱导THP-1细胞转化为巨噬细胞的适宜剂量与时间。在此基础上利用酶联免疫吸附试验(ELISA)检测不同浓度合成脂肽,刺激THP-1细胞分泌细胞因子的含量,并检测CD14抗体预处理THP-1细胞后对细胞因子分泌水平的影响。结果 THP-1细胞在5 ng/ml PMA浓度诱导下48 h分化较为成熟,可以作为体外实验的巨噬细胞模型。TP15、TP17和TP47均可诱导THP-1细胞分泌肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-8。以1 000 ng/ml TP47诱导分泌水平较高。CD14抗体预处理后细胞对3种脂肽的反应能力均下降,所检测的3种细胞因子(TNF-α、IL-1β、IL-8)分泌量均较对照组减少,其中IL-1β及IL-8的分泌浓度下降较为明显。结论 TP15、TP17和TP47均能诱导TPH-1产生促炎性细胞因子,但是TP15和TP17的刺激能力不如TP47。CD14分子参与了TP15、TP17和TP47活化THP-1细胞产生细胞因子的过程。本研究采用根据TP膜脂蛋白N端合成的脂肽,可能不能完全真实反映TP天然膜脂蛋白的活性,需要进一步研究和证实。Objective The study was designated to establish macrophage model and investigate the production of proinflammatory cytokines from macrophages induced by Treponema pallidum lipopeptide. Methods Different doses of PMA were used to induce THP-1 cells for 24 h,36 h and 48 h,then morphology of the cells and the expression of CD14 receptor were observed,so as to determine the appropriate doses and times of PMA intervention. Subsequently,the concentration of TNF-alpha,secreted by THP-1 cell stimulated with TP15,TP17 and TP47,was detected by ELISA,as well as the concentration of the same indicators after blocking the CD14 receptor. Results THP-1 cells induced by 5 ng/ml PMA for 48 h,can be used as a model of macrophage in vitro experiment. TP15,TP17 and TP47 can induce THP-1 cells to secrete TNF-alpha,IL-1beta and IL-8. The level of CKs secreted by THP-1 cells induced by 1000 ng/ml TP47 is higher. The level of IL-1beta and IL-8 decreased notably after blocking CD14 receptor. Conclusions TP15,TP17 and TP47 can induce macrophage to secret cytokines,compared with TP47,the level of cytokines secreted by macrophages stimulated with TP15 and TP17 was lower,which illustrated that The latter two may not be the major pathogenic component of treponema pallidum in syphilis. Three lipopeptides stimulated macrophages through cell surface CD14 receptor molecules,and then lead to the activation of downstream signaling pathways and the secretion of the cytokines.

关 键 词：梅毒螺旋体 脂肽 THP-1 

分 类 号：R759.1[医药卫生—皮肤病学与性病学]