摘要
目的观察气道平滑肌细胞经哮喘致敏血清刺激后嗜酸粒细胞趋化因子(Eotaxin)的表达变化,并探讨其可能的机制。方法贴壁法体外培养大鼠气道平滑肌细胞。建立大鼠哮喘模型,采血收集致敏血清。用致敏血清刺激体外培养的气道平滑肌细胞,或同时予以致敏血清和地塞米松干预。采用酶联免疫吸附法测定培养液上清中Eotaxin水平,放射免疫法测定细胞内环磷酸腺苷(cAMP)表达,逆转录聚合酶链反应半定量检测细胞中Eotaxin mRNA表达。结果致敏血清干预后,细胞培养上清Eotaxin水平和细胞内Eotaxin mRNA表达均较正常对照组显著升高[(107.09±7.12)ng/L比(0.63±0.56)ng/L,P<0.05;1.39±0.04比0.05±0.01,P<0.05],细胞内cAMP表达显著降低[(17.58±3.62)ng/L比(32.39±3.36)ng/L,P<0.05]。地塞米松与致敏血清同时干预后,上清中Eotaxin水平[(64.18±4.04)ng/L]和胞内Eotaxin mRNA(0.77±0.19)表达较哮喘组显著降低,细胞内cAMP表达显著升高[(58.99±5.94)ng/L](P<0.05)。上清Eotaxin水平与胞内cAMP水平呈负相关(r=-0.788,P<0.01)。结论气道平滑肌细胞在致敏状态下Eotaxin基因和蛋白表达均显著增强,可能作为炎性自分泌细胞分泌炎症因子,通过cAMP信号途径参与哮喘的发生。
Objective To observe the eotaxin expression of rat airway smooth muscle cells（ASMCs） induced by serum from asthmatic rats,and explore the possible mechanism.Methods ASMCs isolated from rat tracheas were cultured in vivo.Then they were treated with serum from asthmatic rats,or treated with serum and dexamethasone simultaneously.The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction.The expression of cAMP in ASMCs was examined by radioimmunoassay.Results After the treatment with sensitized serum,the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [（107.09±7.12）ng/L vs.（0.63±0.56）ng/L,P0.05;1.39±0.04 vs.0.05±0.01,P0.05],and the level of cAMP in ASMCs was significantly lower compared with the control group [（17.58±3.62）ng/L vs.（32.39±3.36）ng/L,P0.05].After intervened by the sensitized serum and dexamethasone simultaneously,the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serum alone [（64.18±4.04）ng/L and 0.77±0.19].The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs（r=-0.788,P0.01）.Conclusions There is an autocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum.Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
出处
《中国呼吸与危重监护杂志》
CAS
2010年第5期490-492,共3页
Chinese Journal of Respiratory and Critical Care Medicine
