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

发布时间：2018-06-25 04:08

本文选题：支气管哮喘 + 人软骨糖蛋白　；参考：《中国现代医学杂志》2017年18期

【摘要】：目的探讨小干扰核糖核酸(siRNA)沉默人软骨糖蛋白39(YKL-40)对支气管哮喘小鼠气道平滑肌增殖及迁移的影响。方法 20只健康雌性BALB/c小鼠随机分为健康组(n=10)和哮喘组(n=10),采取腹腔注射抗原和雾化吸入卵蛋的方式复制哮喘模型,健康组小鼠处理与哮喘组相同,只是将致敏物换成生理盐水,留取气管和支气管进行细胞培养,两组小鼠气道壁平滑肌细胞根据转染物不同分为si RNA-YKL-40组、阴性对照组和空白对照组,MTT法检测不同转染组小鼠气道壁平滑肌细胞增殖情况,Transwell法检测不同转染组小鼠气道壁平滑肌细胞迁移能力,实时荧光定量聚合酶链反应(q RT-PCR)和蛋白免疫印迹法(Western blot)检测不同转染组小鼠气道壁平滑肌细胞中YKL-40、白细胞介素4(IL-4)和干扰素-γ(IFN-γ)基因和蛋白表达。结果哮喘组中si RNA-YKL-40组48~96h时细胞吸光度A值均低于阴性对照组和空白对照组,哮喘组中si RNA-YKL-40组、阴性对照组和空白对照组48~96 h时细胞吸光度A值均高于健康组,差异有统计学意义(P0.05);哮喘组中si RNA-YKL-40组迁移细胞数(86.38±8.61)个,低于阴性对照组和空白对照组,哮喘组迁移细胞数均高于健康组,差异有统计学意义(P0.05);哮喘组和健康组中si RNA-YKL-40组YKL-40基因和蛋白相对表达量均低于阴性对照组和空白对照组(P0.05),哮喘组中si RNA-YKL-40组IL-4基因和蛋白相对表达量均低于阴性对照组和空白对照组,而IFN-γ基因和蛋白相对表达量与IFN-γ/IL-4均高于阴性对照组和空白对照组(P0.05),与健康组比较,哮喘组中si RNA-YKL-40组、阴性对照组和空白对照组IL-4基因和蛋白相对表达量均升高,而IFN-γ基因和蛋白相对表达量和IFN-γ/IL-4均降低,差异有统计学意义(P0.05)。结论靶向YKL-40基因沉默能抑制气道壁平滑肌细胞增殖及迁移,可能与调节IL-4/IFN-γ失衡状态而减少气道炎症反应有关。
[Abstract]:Objective to investigate the effect of small interfering ribonucleic acid (siRNA) silencing human cartilage glycoprotein 39 (YKL-40) on airway smooth muscle proliferation and migration in asthmatic mice. Methods Twenty healthy female BALB / c mice were randomly divided into two groups: normal control group (n = 10) and asthmatic group (n = 10). The asthmatic model was induced by intraperitoneal injection of antigens and atomized eggs. The healthy mice were treated the same as the asthmatic group, but the sensitizers were replaced by normal saline. The airway wall smooth muscle cells of the two groups were divided into si RNA-YKL-40 group according to the transfection. The proliferation of smooth muscle cells in airway wall of mice in different transfection groups was detected by MTT assay and the migration ability of smooth muscle cells in airway wall of mice in different transfection groups was detected by Transwell method. The expression of YKL-40, interleukin-4 (IL-4) and interferon- 纬 (IFN- 纬) genes and proteins in airway wall smooth muscle cells of mice in different transfection groups were detected by real-time quantitative polymerase chain reaction (Q RT-PCR) and Western blot (Western blot). Results the absorbance A value of siRNA-YKL-40 group was lower than that of negative control group and blank control group at 4896 h. The absorbance A value of siRNA-YKL-40 group, negative control group and blank control group was higher than that of healthy group at 4896 h. The number of migration cells in the asthma group (86.38 卤8.61) was lower than that in the negative control group and the blank control group, and the number of the migration cells in the asthma group was higher than that in the healthy group. The relative expression of siRNA-YKL-40 gene and protein in asthma group and healthy group was lower than that in negative control group and blank control group (P0.05), and the relative expression of IL-4 gene and protein in siRNA-YKL-40 group was lower than that in negative control group (P0.05). Radiation group and blank control group, The relative expression of IFN- 纬 gene and protein and IFN- 纬 / IL-4 were higher than those of negative control group and blank control group (P0.05). Compared with healthy group, the relative expression of IL-4 gene and protein in asthma group, negative control group and blank control group were increased. The relative expression of IFN- 纬 gene and protein and IFN- 纬 / IL-4 decreased significantly (P0.05). Conclusion targeting YKL-40 gene silencing can inhibit the proliferation and migration of airway smooth muscle cells, which may be related to the regulation of IL-4 / IFN- 纬 imbalance and the reduction of airway inflammation.
【作者单位】：贵州省人民医院胸外科;
【基金】：贵州省科技计划(黔科合SY字,No:20123101)
【分类号】：R562.25

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