中国现代医学杂志2017，Vol.27Issue(18)：15-21,7.DOI:10.3969/j.issn.1005-8982.2017.18.003
siRNA沉默YKL-40对支气管哮喘小鼠气道平滑肌增殖及迁移的影响
Effect of silencing YKL-40 by siRNA on proliferation and migration of airway smooth muscle cells in asthmatic mice
摘要
Abstract
Objective To investigate the effects of silencing human cartilage glycoprotein-39 (YKL-40) by small interfering ribonucleic acid (siRNA) on the proliferation and migration of airway smooth muscle cells in asthmatic mice.Methods Twenty healthy female BALB/c mice were randomly divided into healthy group (n =10) and asthma group (n =10).Asthma models were established by intraperitoneal injection of antigens and inhalation of ovum (OVA).The mice in the healthy group were treated the same as the asthma group,except that the sensitizers were replaced by saline.Tracheal and bronchial tubes were used for cell culture.According to the different transfectants,the airway smooth muscle cells of the two groups were divided into siRNA-YKL-40 group,negative control group and blank control group.The proliferation of airway smooth muscle cells of mice in different transfected groups were detected by MTT assay.The migration abilities of airway smooth muscle cells of mice in different transfected groups were detected by Transwell assay.The expressions of YKL-40,interleukin 4 (IL-4) and interferon-γ (IFN-γ) genes and proteins in airway smooth muscle cells in different transfected groups were detected by qRT-PCR and Western blot,respectively.Results In the asthma group,the absorbance A values at 48,72 and 96 h of the siRNA-YKL-40 group were significantly lower than the negative control group and blank control group and the absorbance A values at 48,72 and 96 h of the siRNA-YKL-40 group,negative control group and blank control group in the asthma group were significantly higher than the healthy group,(P ＜ 0.05).In the asthma group,the number of migrated cells of the siRNA-YKL-40 group was (86.38± 8.61),which was significantly lower than the negative control group and blank control group and the number of migrating cells of the asthma group was significantly higher than the healthy group (P＜ 0.05).In the asthma group and the healthy group,the relative expression levels of YKL-40 gene and protein in the siRNA-YKL-40 group were lower than the negative control group and blank control group (P＜ 0.05),in the asthma group,the relative expression levels of IL-4 gene and protein in the siRNA-YKL-40 group were lower than the negative control group and blank control group,while the relative expression levels of IFN-γ gene and protein and the value of IFN-γ/IL-4 were significantly higher than the negative control group and blank control group (P ＜ 0.05).Compared with the healthy group,the relative expression levels of IL-4 gene and protein in the siRNA-YKL-40 group,negative control group and blank control group in the asthmatic group were significantly increased,while the relative expression levels of IFN-γ gene and protein and the value of IFN-γ/IL-4 were significantly decreased (P ＜ 0.05).Conclusions Gene silencing targeting YKL-40 inhibits airway smooth muscle cell proliferation and migration.It might be related to the regulation of the imbalance of IL-4/IFN-γ to reduce airway inflammation.关键词
支气管哮喘/人软骨糖蛋白39/沉默核糖核酸/细胞增殖/细胞迁移Key words
bronchial asthma/human cartilage glycoprotein-39/RNA silencing/cell proliferation/cell migration分类
医药卫生引用本文复制引用
韩连奎,许川,梅宏,孙勇攀,刘波..siRNA沉默YKL-40对支气管哮喘小鼠气道平滑肌增殖及迁移的影响[J].中国现代医学杂志,2017,27(18):15-21,7.基金项目
贵州省科技计划(黔科合SY字,No:20123101) （黔科合SY字,No:20123101）
