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目的探讨IL-22对人气道上皮细胞、气道平滑肌细胞、气道成纤维细胞的生理学作用。方法用实时定量PCR检测气道上皮细胞、气道平滑肌细胞、气道成纤维细胞哮喘血清刺激前后IL-22R1 mRNA表达的变化。不同浓度的IL-22(10 ng/ml,100 ng/ml,1 000 ng/ml)刺激气道上皮细胞、气道平滑肌细胞和气道成纤维细胞后,用MTT法检测细胞的增殖,用流式细胞技术(FACS)检测细胞的凋亡和坏死。结果哮喘血清刺激后气道上皮细胞IL-22R1 mRNA表达降至正常对照组的9%,气道平滑肌细胞IL-22R1 mRNA表达升高至正常对照组的345倍,而气道成纤维细胞IL-22R1 mRNA表达无明显变化。高剂量(1 000 ng/ml)的IL-22刺激气道上皮细胞和气道成纤维细胞12、24 h可显著降低细胞增殖(P<0.05,P<0.01)。三种不同浓度的IL-22刺激气道上皮细胞24 h后均导致细胞凋亡显著下降(P<0.05),低浓度的IL-22(10 ng/ml)导致细胞坏死增加(P<0.01)。不同浓度的IL-22刺激气道平滑肌细胞后,细胞凋亡和坏死无显著性变化。中、高浓度IL-22(100 ng/ml,1 000 ng/ml)刺激气道成纤维细胞后,细胞坏死率显著下降(P<0.05)。结论 IL-22在支气管哮喘中对气道上皮的作用具有双重性,并与支气管哮喘的病程相关;而对气道平滑肌细胞的作用则可能与浓度及病程相关。支气管哮喘后续阶段,IL-22对气道成纤维细胞作用轻微。
Objective To investigate the physiological effects of IL-22 on human airway epithelial cells, airway smooth muscle cells and airway fibroblasts. Methods Real-time quantitative PCR was used to detect the expression of IL-22R1 mRNA in airway epithelial cells, airway smooth muscle cells and airway fibroblasts before and after stimulation. After stimulating airway epithelial cells, airway smooth muscle cells and airway fibroblasts with different concentrations of IL-22 (10 ng / ml, 100 ng / ml, 1000 ng / ml), the cell proliferation was measured by MTT assay, Cell Technology (FACS) detects apoptosis and necrosis of cells. Results The expression of IL-22R1 mRNA in airway epithelial cells was decreased to 9% in normal control group and IL-22R1 mRNA in airway smooth muscle cells was 345 times higher than that in normal control group, while IL-22R1 in airway fibroblasts No significant change in mRNA expression. High dose (1 000 ng / ml) of IL-22 stimulated airway epithelial cells and airway fibroblasts for 12,24 h, which could significantly reduce cell proliferation (P <0.05, P <0.01). Stimulation of airway epithelial cells with three concentrations of IL-22 for 24 h resulted in a significant decrease of apoptosis (P <0.05). Low concentrations of IL-22 (10 ng / ml) resulted in an increase of cell necrosis (P <0.01) . After IL-22 stimulation of airway smooth muscle cells with different concentrations, there was no significant change in apoptosis and necrosis. The cell necrosis rate was significantly decreased after medium and high concentrations of IL-22 (100 ng / ml, 1 000 ng / ml) stimulated airway fibroblasts (P <0.05). Conclusions IL-22 plays a dual role in airway epithelium in bronchial asthma and is related to the course of bronchial asthma. The effect of IL-22 on airway smooth muscle cells may be related to the concentration and course of disease. In the subsequent stages of bronchial asthma, IL-22 has a slight effect on airway fibroblasts.