| 摘要: |
| 目的:探讨烟曲霉菌对呼吸道上皮细胞的粒-巨噬细胞集落刺激因子(GM-CSF)表达的影响及其可能机制。方法:分别应用不同浓度(0、8、16、20 mg/L) 的烟曲霉菌提取物(Aspergillus fumigatus extract,AFE)作用体外培养人支气管上皮细胞16HBE-14o不同的时间(12、24、48、72 h)。同时还应用热处理的AFE(heat-treated AFE,HT-AFE)、丝氨酸蛋白酶抑制剂(抑肽酶)、蛋白激酶激活受体-2(PAR-2)激动剂(SLIGLV- NH2)和PAR-2阻断剂(FSLLRY-NH2)作用16HBE-14o细胞。应用ELISA法检测细胞上清液GM-CSF含量,同时应用RT-PCR法检测GM-CSF mRNA 的表达。结果:正常培养条件下的对照组细胞仅表达分泌少量GM-CSF,而给予不同浓度的AFE作用后,细胞合成和分泌GM-CSF的量较对照组明显增加(P<0.05),并呈明显的时间和浓度依赖性。PAR-2激动剂同样可以促进细胞合成分泌GM-CSF。丝氨酸蛋白酶抑制剂和PAR-2阻断剂几乎完全抑制AFE的上述作用。HT-AFE的蛋白酶活性完全灭活,不能促进细胞GM-CSF表达增加。结论:AFE依赖其蛋白酶活性激活PAR-2受体,促进呼吸道上皮细胞GM-CSF合成和分泌,从而促进哮喘的发生和发展。 |
| 关键词: 烟曲霉菌 支气管上皮细胞 粒-巨噬细胞集落刺激因子 蛋白酶激活受体-2 |
| DOI：10.3724/SP.J.1008.2008.00917 |
| 投稿时间：2008-01-08修订日期：2008-02-28 |
| 基金项目: |
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| Effects of Aspergillus fumigatus extract on expression of GM-CSF in human bronchial epithelial cells |
| LONG Fei,GAO Fu-sheng,DING Xing,QI Hui-hui,JIN Xian-qiao* |
| (Department of Respiratory Medicine,First Peoples’ Hospital of Shanghai,Shanghai Jiaotong University,Shanghai 200080,China) |
| Abstract: |
| Objective:To investigate the effects of Aspergillus fumigatus extract (AFE) on the expression of GM-CSF in human bronchial epithelial cells and its possible mechanism.Methods: Human bronchial epithelial cells (16HBE-14o) were cultured in vitro and were exposed to different concentrations of AFE (0,8,16 and 20 mg/L) for different peroids (12、24、48、72 h).Moreover,heat-treated AFE,serine protease inhibitors (aprotinin),protease-activated receptor-2 (PAR-2) agonist (SLIGLV- NH2) and PAR-2 antagonist (FSLLRY-NH2) were also used to treat 16HBE-14o cells.The production and release of GM-CSF in different supernatants was determined by ELISA.The expression of GM-CSF mRNA was measured by RT-PCR.Results: 16HBE-14o cells in the normal control group only had slight expression of GM-CSF; the expression was significantly increased after AFE treatment(P<0.01); and the effect of AFE was in a time- and dose-dependent manner.PAR-2 agonist also promoted release of GM-CSF.Aprotinin and PAR-2 antagonist (FSLLRY-NH2) almost totally inhibited the effect of AFE on GM-CSF production by 16HBE-14o cells.Heat-treated AFE,which lost protease activities,exerted no effect on GM-CSF production.Conclusion: AFE,with its protease activity,can activate PAR-2,and subsequently causes GM-CSF synthesis and release by airway epithelial cells,which may contribute to the development and deterioration of asthma. |
| Key words: Aspergillus fumigatus bronchial epithelial cell granulocyte-macrophage colony stimulating factor protease-activated receptor-2 |
