关键词: 哮喘, 肥大细胞, 反义寡核苷酸, 干细胞因子, 成纤维细胞

Abstract: Objective To reveal the role of stem cell factor(SCF)- mediating fibroblasts and mast cells interaction in pathophysiology process of asthma. Methods We transfected NIH3T3 cells with SCF antisense oligonucleotide(SCF ASON) and detected SCF expression by immunochemistry and RT-PCR. Then, we separated mast cells from mice bone marrow and established NIH3T3 and mast cells cocultures. After 10μg/mL SCF ASON intervention, histamine and eotaxin levels in culture supernatants were determined by ELISA and fluorometry. Growth curves of fibroblasts and mast cells were drawn. We also observed mast cells apoptosis by AO stain and flow cytometric analysis. Results SCF ASON strikingly down-regulated SCF protein and mRNA level of NIH3T3 cells. SCF ASON intervention can inhibit the growth of NIH3T3，and induce mast cell apoptosis in cocultures（14.0%±0.81% at 96 hour）. Both histamine（3.08±0.38ng/ml vs 3.83±0.41ng/ml, P<0.05）and eotaxin ( 4.398±0.333 vs 5.785±0.395ng/ml, P<0.05 )decreased significantly. Conclusions SCF may take part in the pathophysiology process of asthma, probably by inhibiting apoptosis of mast cells, contributing to mast cells activation and fibroblast proliferation.

Key words: asthma, mast cells, antisense oligonucleotide, stem cell factor, fibroblast