sCD14在内毒素诱导巨噬细胞活化中的调节机制

目的 观察不同浓度的sCD14和脂多糖（LPS）经过孵育和不孵育同时刺激佛波醇酯（PMA）诱导分化的THP-1细胞分泌肿瘤坏死因子-α（TNF-α）,以研究sCD14在LPS转运和活化过程中的重要调节作用.方法 通过细胞培养,实验细胞为PMA诱导分化的THP-1细胞;同时将实验分为sCD14组（终浓度分别为1、0.5、0.1、0.01μg/ml）、sCD14～LPS混合物预孵育组（LPS终浓度分别为10、1ng/ml;sCD14终浓度为分别1、0.5、0.1、0.01μg/m1）、sCD14-LPS不孵育组（LPS终浓度分别为10、1ng/ml;sCD14终浓度为1、0.5、0.1、0.01μg/ml）,并分别培养4h后,吸出上清液至1.5ml的离心管中,于-70℃内保存并采用化学发光法检测TNF-α.结果 sCD14本身不能刺激单核细胞分泌TNF-α;LPS浓度为1ng/ml、sCD14浓度为0.01μg/ml和0.1μg/ml时,与LPS单独刺激TNF-α分泌量的差异均无统计学意义（均P＞0.05）;sCD14浓度为0.5μg/ml和1μg/ml时,TNF-α分泌量的差异均有统计学意义（P＜0.05或0.01）.在LPS浓度为10ng/ml、sCD14浓度为0.5μg/ml和1μg/ml时,与LPS单独作用TNF-α分泌量均有显著性差异（均P＜0.01）.sCD14/LPS混合物预孵育组曲线低平,无明显分泌高峰.结论 适当浓度的sCD14可明显增强LPS对单核巨噬细胞的活化作用,且在LPS高浓度时尤为明显,而将sCD14和LPS混合孵育可使上述效应减少甚至消失.

Regulatory role of sCD14 in macrophage activation and cytokines secretion stimulated by LPS

Objective To investigate the regulatory role of sCD14 in macrophage activation and cytokine secretion stimulated by lipopolysaccharide （LPS）.Methods THP-1 cells were differentiated by PMA, and divided into three groups： sCD14 group （final concentration： 1, 0.5, 0. 1 and 0.01 μ g/mL）, sCD14/LPS pre-incubation group （LPS fina） concentration： 10 and lng/mL; sCD14 final concentration： 1, 0.5, 0.1 and 0.01μg/mL）, sCD14/LPS without preincubation （final concentration of LPS and sCD14 were as above）. The cells were cultured for 4h and the TNF- αlevels in supernatant were measured.Results sCD14 did not stimulate mononucle cells to secrete TNF-α. Low concentration sCD14 （0.01 and 0.1 μ g/mL） did not increase TNF-α secretion when cells were stimulate by lng/mL LPS（P 〉0.05）. High concentration sCD14 （0.5 and 1 μg/mL） increased TNF- α secretion（P 〈0.05）; the effect was more prominent when LPS concentration was 10ng/mL （P 〈0.01 ）. Conclusion sCD14 can significantly enhance LPS activation on monocyte-macrophage.