PMA对THP-1细胞和RA患者单核细胞及培养上清中CD147表达的影响

目的：研究豆蔻佛波醇乙酯（PMA）刺激前后,体外培养的人单核细胞系THP-1细胞和类风湿关节炎（RA）患者外周血单核细胞（human peripheral blood monocytes,HPBM）上及培养上清中CD147的表达.方法：以贴壁法分离RA患者HPBM.用PMA刺激体外培养的RA患者HPBM和THP-1细胞,采用流式细胞术动态检测THP-1细胞、RA患者HPBM膜表面CD147的表达.用双抗体夹心ELISA法检测培养上清中CD147分子的含量.结果：PMA刺激前,THP-1细胞膜及RA患者HPBM表面CD147分子的表达均较高,培养上清中均可检测到CD147分子.PMA刺激后,THP-1细胞培养上清中CD147的含量升高,THP-1细胞上CD147的表达先升高后降低,最后进入稳定期;RA患者的HPBM上CD147的表达下降,培养上清中CD147的含量升高,于刺激后2 d进入稳定期.结论：THP-1细胞、RA患者HPBM膜表面CD147分子可从细胞上脱落或由细胞直接分泌到培养液中.PMA可上调可溶性CD147的表达.

Effects of PMA on CD147 expression in cultured supernatants of in vitro cultured THP-1 cells and on monocytes of RA patients

AIM: To investigate the expression of CD147 in culture supernatant of in vitro cultured THP-1 cells and on monocytes of RA patients before and after stimulation with phorbol myristate acetate(PMA). METHODS: Human peripheral blood monocytes (HPBMs) were segregated from RA patients by adherent culture method. After the HPBMs and THP-1 cells were stimulated with PMA, the expression rate and expression intensity of CD147 on THP-1 cells and HPBMs of RA patients were determined by immunofluorescence flow cytometry. The soluble CD147 levels in culture supernatant of in vitro cultured THP-1 cells and HPBMs were quantified by double antibody sandwich ELISA. RESULTS: The soluble CD147 in the supernatant of cultured THP-1 cells and HPBMs was detected before PMA stimulation.Both cells showed high CD147 expression. After PMA stimulation, soluble CD147 level in the supernatant of THP-1 cells increased. CD147 expression on THP-1 cells first increased then decreased and kept stable several days later. CD147 expression on HPBMs of RA patients decreased, but CD147 level in the culture supernatant of HPBMs of RA patients increased and kept stable 2 days later. CONCLUSION: CD147 molecule can shed from the membrane of THP-1 cells and HPBMs of RA patients, or it can be directly secreted into the culture fluid from the two cells. PMA can promote expression of soluble CD147.