烟曲霉菌提取物对人支气管上皮细胞粒-巨噬细胞集落刺激因子表达的影响  

Effects of Aspergillus fumigatus extract on expression of GM-CSF in human bronchial epithelial cells

作　　者：龙飞[1] 高福生[1] 丁星[1] 祁卉卉[1] 金先桥[1] 

机构地区：[1]上海交通大学附属第一人民医院呼吸内科,上海200080

出　　处：《第二军医大学学报》2008年第8期917-920,共4页Academic Journal of Second Military Medical University

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。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-aetivated 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.

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 

分 类 号：R363[医药卫生—病理学]