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目的观察吸入型糖皮质激素二丙酸倍氯米松(BDP)和布地奈德(BUD)对白细胞介素13(IL-13)激活的肺成纤维细胞的影响。方法 IL-13加入培养的肺成纤维细胞,采用噻唑蓝法、免疫组织化学法、免疫印迹法、逆转录聚合酶链反应、ELISA 等方法检测细胞增殖、α-肌动蛋白(α-SMA)表达及分泌白细胞介素6(IL-6)、嗜酸粒细胞趋化因子(eotaxin)的变化,观察 BDP 和 BUD 的作用。结果加入20 ng/ml 的 IL-13后,成纤维细胞分泌 IL-6明显增加[(20±2)ng/L和(140±8)ng/L],eotaxin 也明显增加[(64±25)ng/L 和(334±51)ng/L]。10~(-8)mol/L～10~(-5)mol/L 的 BDP 组和 BUD组 IL-6表达水平[(112±3)ng/L～(53±2)ng/L 和(55±14)ng/L～(32±6)ng/L]比 IL-13组明显下降,IL-6 mRNA 表达比 IL-13组明显减少,细胞上清液中 eotaxin 水平[(297±59)～(226±43)ng/L和(287±59)～(183±43)ng/L]比 IL-13组明显降低。IL-13诱导成纤维细胞表达α-SMA mRNA 及蛋白明显增加,并促进成纤维细胞增殖(1.5±0.2)倍。BDP 组和 BUD 组对 IL-13诱导的α-SMAmRNA 及蛋白的表达无显著影响,并明显具有协同 IL-13促进成纤维细胞增殖的作用。结论 BDP和 BUD 对 IL-13刺激成纤维细胞的作用具有多重调节效应。BDP 和 BUD 抑制 IL-13诱导成纤维细胞合成、释放炎症介质 IL-6和 eotaxin 的作用,有利于改善气道上皮下纤维化,但对于 IL-13促进成纤维细胞转化为肌成纤维细胞及其增殖方面起负面影响。
Objective To observe the effects of inhaled glucocorticoid beclometasone dipropionate (BDP) and budesonide (BUD) on interleukin-13 (IL-13) activated pulmonary fibroblasts. Methods IL-13 was added into cultured lung fibroblasts. The cell proliferation, α-actin (α-SMA) and cell proliferation were measured by using thiazolyl blue method, immunohistochemistry, Western blotting, reverse transcription polymerase chain reaction Expression and secretion of interleukin-6 (IL-6), eotaxin changes observed BDP and BUD role. Results The IL-6 secretion of fibroblasts was significantly increased by 20 ng / ml IL-13 [(20 ± 2) ng / L and (140 ± 8) ng / L] ) ng / L and (334 ± 51) ng / L]. The levels of IL-6 in BDP group and BUD group [(112 ± 3) ng / L ~ (53 ± 2) ng / L and 55 ± 14 ng / L 32 ± 6 ng / L] significantly decreased IL-6 mRNA expression compared with IL-13 group, and the level of eotaxin in the cell supernatant [(297 ± 59 ) ~ (226 ± 43) ng / L and (287 ± 59) ~ (183 ± 43) ng / L] were significantly lower than those in IL-13 group. IL-13 induced a significant increase ofα-SMA mRNA and protein expression in fibroblasts and promoted fibroblast proliferation (1.5 ± 0.2) fold. BDP group and BUD group had no significant effect on the expression of α-SMA mRNA and protein induced by IL-13 and obviously had the synergistic effect of IL-13 on fibroblast proliferation. Conclusion BDP and BUD have multiple regulatory effects on IL-13-stimulated fibroblasts. BDP and BUD can inhibit the synthesis of IL-13-induced fibroblasts and release the inflammatory mediators IL-6 and eotaxin, which can help to improve airway subcutaneous fibrosis. However, IL-13 promotes the transformation of fibroblasts into myofibroblasts and their Proliferation negatively affected.