咯利普兰对佐剂关节炎大鼠腹腔巨噬细胞体外释放炎性细胞因子的影响

Effects of rolipram on release of inflammatory cytokines in vitro by peritoneal macrophages extracted from adjuvant arthritis rats

目的探讨选择性磷酸二酯酶(PDE)4抑制剂咯利普兰对脂多糖(LPS)诱导的佐剂关节炎(AA)大鼠腹腔巨噬细胞(PM)体外释放促炎细胞因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的影响,为选择性PDE4抑制剂治疗类风湿关节炎(RA)提供理论依据。方法制备AA大鼠模型,收集PM,体外给予LPS及不同浓度咯利普兰共同孵育,酶联免疫吸附测定法(ELISA)检测细胞培养上清中TNF-α、IL-1β水平,反转录聚合酶链反应(RT-PCR)法测定细胞内TNF-αmRNA、IL-1βmRNA的表达,并与正常对照相比较。结果 AA大鼠PM培养上清中TNF-α、IL-1β、细胞内TNF-αmRNA、IL-1βmRNA表达均高于对照组大鼠,分别为(386.80±63.24)ng/L vs(48.53±6.47)ng/L(P<0.01)、(317.77±29.33)ng/L vs(77.75±11.60)ng/L(P<0.01)、77.92±12.55vs 15.97±4.37(P<0.01)、49.30±10.06vs 10.15±1.34(P<0.01)。咯利普兰能够剂量依赖性地抑制LPS刺激的AA大鼠PM体外表达TNF-αmRNA、IL-1βmRNA,并抑制TNF-α、IL-1β产生。结论咯利普兰能抑制LPS诱导的AA大鼠PM体外释放促炎性细胞因子,提示其可能通过抑制炎症反应用于RA的治疗。

Objective To explore the effects of selective phosphodiesterase（PDE） 4 inhibitor rolipram on lipopolysaccharide（LPS）-induced releasing of proinflammatory cytokines tumor necrosis factor alpha（TNF-α）,interleukin 1 beta（IL-1β） in vitro by peritoneal macrophages（PM） extracted from adjuvant arthritis（AA） rats.Methods AA rat model and normal control were prepared,then PM were collected and cultured in vitro,LPS was administrated and different concentrations of rolipram were added at the same time.Levels of TNF-α,IL-1β in cell culture supernatant were assayed by enzyme-linked immunosorbent assay（ELISA）,and intracellular expression of TNF-α mRNA,IL-1β mRNA were determined by reverse transcription-polymerase chain reaction（RT-PCR）.Results TNF-α,IL-1β in PM culture supernatants,intracellular expression of TNF-α mRNA,IL-1β mRNA of AA rats were significantly higher than those of normal rats,（386.80±63.24） ng/L vs（48.53±6.47） ng/L（P0.01）,（317.77±29.33） ng/L vs（77.75±11.60） ng/L（P0.01）,77.92±12.55 vs 15.97±4.37（P0.01）,49.30±10.06 vs 10.15±1.34（P0.01） respectively.Rolipram in a dose-dependent manner inhibited the LPS-stimulated expression of TNF-α mRNA,IL-1β mRNA,and inhibited TNF-α,IL-1β production in AA rats.Conclusion Rolipram can inhibit LPS-induced releasing of pro-inflammatory cytokines ex vitro by PM extracted from AA rats,suggested that the selective PDE4 inhibitors may be used in the treatment of RA by inhibiting the inflammatory reaction.