辛伐他汀对单个核细胞合成白细胞介素-17及转录因子B细胞活化转录因子的影响

Effect of simvastatin on interleukin-17 production and expression of interleukin-17 transcription factor B-cell activating transcription factor in peripheral blood mononuclear cells from rheumatoid arthritis patients and healthy individuals

目的研究辛伐他汀对类风湿关节炎（RA）患者外周血单个核细胞（PBMCs）合成和分泌白细胞介素（IL）-17的作用及对B细胞活化转录因子（BATF）的影响。方法健康人和RA患者PBMCs在有或无辛伐他汀，甲羟戊酸作用下，予以佛波酯和离子霉素共刺激培养。采用实时-聚合酶链反应（RT-PCR）技术分析IL-17mRNA和BATFmRNA表达水平，以3-磷酸甘油醛脱氢酶（GAPDH）为内参；采用酶联免疫吸附试验（ELISA）检测各组细胞培养液中IL-17浓度；采用蛋白印迹法分析BATF蛋白在辛伐他汀作用后表达的变化。2组比较采用配对t检验，多组间比较采用方差分析。结果RA患者[（79．6±12．7）与（15．8±5．8）pg/ml，P〈0．05；（90．3±9．7）与（12．9±7．9）pg／m1，P〈0．05]和健康者[（69．2±12．2）与（8．1±2．2）pg／ml，P〈0．05；（76．6±14．7）与（10．2±7．2）pg／ml，P〈0．05]PBMCs经佛波酯和离子霉素激活48h或72h后IL-17水平显著升高。加入辛伐他汀作用后，20μmol/L辛伐他汀较10μmol/L辛伐他汀更明显地抑制了佛波酯和离子霉素所诱导IL-17的基因转录和蛋白合成。甲羟戊酸与辛伐他汀共同作用于PBMCs后，IL-17产生水平升高不明显。辛伐他汀作用后BATF水平无明显减弱。结论辛伐他汀抑制外源刺激下PBMCs合成和分泌IL-17，这种作用并不能通过抑制转录因子BATF的水平而发挥作用。

Objective To investigate the effects of simvastatin on the production of intedeukin （IL）-17 and B-cell activating transcription factor （BATF） in the peripheral blood mononuelear cells （PBMCs） of rheumatoid arthritis （RA） patients and healthy individuals. Methods PBMCs were isolated from heparinized blood of healthy donors or RA patients using Ficoll-Hypaque density gradient centrifugation. The cells were stimulated by PMA and ionomycin in the absence or presence of simvastatin or MVA at 37 ℃ 5%CO2. The reRAN level of IL-17, BATF and GAPDH was detected by RT-PCR; the protein level of IL-17 in supematants was assayed by ELISA kit; and the protein level of BATF was detected by Western Blotting. The comparison between the two groups was carried out by paired-t test and Chi-square test was used for multi-group comparison. Results PBMCs of healthy donors [（69.2±12.2） vs （8.1±2.2） pg/ml, P〈0.05; （76.6±14.7） vs （10.2±7.2） pg/ml, P〈0.05] and RA patients [（79.6_±12.7） vs （15.8±5.8） pg/rnl, P〈0.05; （90.3±9.7） vs （12.9±7.9） pg/ml, P〈0.05] were stimulated with PMA and ionomycin to produce high levels of IL-17. After treatment with simvastatin, the expression and secretion level of IL-17 in healthy controls and RA PBMCs were markedly decreased. The inhibition of simvastatin on the production of IL-17 was reversed by mcvalonic acid （MVA）, but no significant changes of BATF after treating with simvastatin. Conclusion Simvastatin inhibits the production of IL-17 in the PBMCs at gene and protein levels, which is not targeted at suppressing the expression of IL-17 transcription factor BATF.