PI3K在细胞外基质分泌细胞因子表达中的作用  

The role of PI3K in regulating cytokine secretion by extracellular matrixes

作　　者：刘瑾[1] 施珊娜[1] 许淑云[1] 徐永健[1] 胥杰[1] 陈忠仁[1] 王正艳[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸内科中国卫生部呼吸系疾病重点实验室,武汉430030

出　　处：《中华微生物学和免疫学杂志》2009年第7期597-601,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30500223）;湖北省自然科学基金资助项目（2007ABA157）

摘　　要：目的探讨纤维连接蛋白（FN）、Ⅰ型胶原蛋白（ColⅠ）两种细胞外基质（ECM）成分对被动致敏的人气道平滑肌细胞（HASMCs）免疫功能的影响及磷脂酰肌醇-3激酶（P13K）在此调控中的作用。方法将HASMCs接种于FN、ColⅠ包被的培养板和空白培养板中，用10％哮喘患者血清被动致敏HASMCs，以10％非哮喘者血清为对照，在加入血清前用PBK抑制剂（LY294002）预处理HASMCs 30min。采用RT—PCR法检测HASMCs RANTES（regulated upon activation，normal T cell expressed and secreted）、Eotaxin、TGF—β1 mRNA表达；ELISA法测定HASMCs培养上清中RANTES、Eotaxin、TGF—β1蛋白水平。结果与单纯对照血清组比较，单纯哮喘血清组、对照血清＋FN组、对照血清＋ColⅠ组HASMCs RANTES、Eotaxin、TGF-β1 mRNA和HASMCs培养上清中蛋白的表达均增高（P〈0．05）。哮喘血清＋FN、哮喘血清＋ColⅠ处理HASMC后，RANTES、Eotaxin、TGF—β1 mRNA和HASMCs培养上清中蛋白的表达均高于单纯哮喘血清组，且LY294002干预后上述各指标均下降（P〈0．05）。结论细胞外基质对被动致敏的HASMCs免疫功能具有调控作用，PI3K可能参与细胞外基质对被动致敏的HASMCs的免疫功能调控。Objective To investigate the regulatory effects of extracellular matrixes, including fibronectin（FN） , and collagen Ⅰ （ Col Ⅰ ） on the immunologic function of human airway smooth muscle cells （HASMCs） passively sensitized with asthmatic serum, and the role of phosphoinositide 3-kinase （ PI3 K）. Methods Primarily cultured HASMCs were inoculated on the blank plates or on the plates coated with difference matrix proteins, added 10% asthmatic serum to passively sensitized non-asthmatic HASMCs and 10% non-asthmatic serum treated HASMCs as control, cell pretreated with PI3K inhibitor LY294002 for 30 min. The expressions of RANTES, Eotaxin, TGF-β1 mRNA were observed by RT-PCR and RANTES, Eotaxin, TGF-β1 protein in the cell culture supernatants was detected by enzyme-linked immunosorbent assay （ELISA）. Results Compared with the control serum group, the expressions of RANTES, Eotaxin, TGF- β1 mRNA of HASMCs and those protein in HASMCs culture supernatants were significantly increased in the asthmatic serum group and the control serum ＋ FN group and the control serum ＋ Col Ⅰ group （ P 〈 0.05 ）. The expressions of RANTES, Eotaxin, TGF-β1 mRNA of HASMCs and protein in HASMCs culture supernatants were significantly increased in the asthmatic serum ＋ FN group and the asthmatic serum ＋ Col Ⅰ group. 50 μmol/L LY294002 could significantly inhibit the expressions of RANTES, Eotaxin, TGF-β1 mRNA of HASMCs and protein in HASMCs culture supernatants（P 〈 0.05 ）. Conclusion These results suggest extracellular matrixe may regulate immunomodulatory function of HASMCs passively sensitized with asthmatic serum and PI3K signaling pathway may play an important role in the process.

关 键 词：哮喘 细胞外基质 气道平滑肌细胞 免疫功能 磷脂酰肌醇-3激酶 

分 类 号：R562.25[医药卫生—呼吸系统]