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目的研究免疫调节剂咪喹莫特对卵蛋白(OVA)致敏大鼠支气管旁淋巴结(PBLN)细胞培养体系中T辅助淋巴细胞(Th)亚群产生细胞因子的影响,并探讨其作用机制。方法建立PBLN细胞培养体系,并按不同浓度分为A～F组进行干预。在培养0、3、6、12、24、48h后,各组分别取5孔细胞培养液,用酶联免疫吸附法(ELISA)检测各细胞因子的蛋白质水平,用逆转录聚合酶链反应(RTPCR)检测细胞沉淀中白细胞介素(IL)4和干扰素(IFN)γ等细胞因子的mRNA表达。结果在A组的各时间点PBLN细胞培养液中,仅可检测到少量IFNγ。随着培养时间的延长,咪喹莫特浓度为1、10μg/ml的E和F组与B组相比,IL4及相关mRNA水平增加缓慢,IFNγ水平增长迅速(均P<0.005)。此作用从培养6h开始,12h达峰值,持续至24h。在培养12～48h时,C组IL4表达与B组比,差异有统计学意义(P<0.05),而IFNγ表达无差异(P>0.05)。结论咪喹莫特对致敏大鼠PBLN细胞培养体系中Th亚群产生细胞因子的最佳作用发生在12h时,提示咪喹莫特可能在支气管哮喘等特异性疾病的迟发炎症反应阶段发挥重要作用。
Objective To study the effect of imiquimod, an immunomodulator, on the production of cytokines by T helper lymphocyte (Th) subsets in the culture system of ovalbumin (OVA) -induced bronchogenic lymph nodes (PBLN) in rats and to explore its possible mechanism. Methods PBLN cell culture system was established and divided into A ~ F groups according to different concentrations. After culturing for 0, 3, 6, 12, 24, and 48h, 5-well culture medium was taken in each group. The protein level of each cytokine was detected by enzyme-linked immunosorbent assay (ELISA) RTPCR) was used to detect the mRNA expression of cytokines such as interleukin (IL) 4 and interferon gamma in cell pellet. As a result, only a small amount of IFNγ was detected in the PBLN cell culture fluid at each time point in group A. Compared with group B, IL4 and related mRNA levels increased slowly and IFNγ levels increased rapidly (P <0.005 for both). This effect from the beginning of culture 6h, peaked at 12h, lasted until 24h. Compared with group B, the expression of IL4 in group C was significantly higher than that in group B (P <0.05), while there was no difference in IFNγ expression (P> 0.05). Conclusion The best effect of imiquimod on cytokine production by Th group in PBLN cell culture system in sensitized rats occurred at 12h, suggesting that imiquimod might play a role in the late inflammatory reaction stage of specific diseases such as bronchial asthma. Important role.