摘要
为了研究RNA干扰基因表达对哮喘小鼠支气管上皮细胞凋亡的影响,本研究拟通过RNA干扰技术降低Toll样受体表达,检测其是否对缓解哮喘小鼠的支气管上皮细胞凋亡发挥作用。本实验将30只小鼠,雌雄各半,随机分为对照组、哮喘组以及干预组3组。哮喘造模成功后,原代培养各组小鼠的支气管上皮细胞,4~8代后进行si RNA干扰。通过RT-PCR以及TUNEL的方法分别检测小鼠细胞内Toll样受体的基因表达量和细胞凋亡情况。结果显示,哮喘的小鼠Toll样受体的表达明显升高,支气管上皮细胞凋亡比例也比较高,与对照组小鼠进行比较,两种指标均有统计学差异;而si RNA进行干扰后,相较于哮喘组,Toll样受体的表达量明显下降;且凋亡细胞的比例也明显下降,与哮喘组进行比较,两组间有明显的统计学差异。本研究在小鼠模型中验证了用RNA干扰技术对Toll样受体基因表达的干预作用,并且也证明此技术能明显改善哮喘小鼠的病理表型,是非常有前景的临床治疗方法,为临床推广提供了较为可靠的基础实验证据。
To study the effect of RNA interference gene expression on apoptosis of epithelial cell in asthmatic mice,this study was designed to reduce the expression of Toll like receptors by RNA interference,and to investigate whether it could alleviate the apoptosis of bronchial epithelial cells in asthmatic mice.In this study,30 mice,half male and half female,were randomly divided into control group,asthma group and intervention group,ten for each group.After asthma disease models were constructed successfully,bronchial epithelial cells in each mice group were first primarily cultured and then its 4~8 generations were cultured with si RNA interference.RT-PCR and TUNEL methods were applied to detect Toll-like receptor gene expression levels and cell apoptosis rate,respectively.The results showed that Toll-like receptors expression was increased significantly in asthmatic mice and the bronchial epithelial apoptosis ratio was higher compared with the control.The differences of the two indexes were statistically significant.After si RNA interference,Toll-like receptors expression was significantly decreased compared with the asthma group.Besides,proportion of apoptosis cells was also significantly decreased compared with asthma group,and there were significant statistical differences between the two groups.The study demonstrated the intervention of RNA interference in the expression of Toll-like receptor genes in mouse models,and also demonstrated that this technique could significantly improve the pathological phenotype in asthmaticmice.It might be a very promising method for clinical treatment and could provide a reliable basis for clinical trials.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第9期3516-3520,共5页
Genomics and Applied Biology
基金
湘南学院
武汉大学人民医院资助
