博来霉素诱导肺纤维化小鼠肺组织炎性microRNA表达的筛选研究

Expression of inflammatory microRNAs in lung of mice with pulmonary fibrosis induced by bleomycin

目的运用PCR Array技术检测肺纤维化模型小鼠肺组织中炎性microRNA(miRNA)表达谱,以炎症为切入点分析miRNA在肺纤维化中可能的作用机制。方法将12只健康雄性SPF级C57BL/6小鼠随机分为对照组和模型组,每组6只。模型组按5 mg/kg体重剂量以滴鼻方式给予博来霉素,对照组给予等量生理盐水。给药14天后处死全部小鼠,提取肺组织总RNA,采用PCR Array技术筛选出炎性miRNA差异表达谱,运用生物信息学知识对差异miRNA进行靶基因预测和富集性分析。结果肺纤维化模型构建成功;与对照组相比,模型组小鼠肺组织中共筛选得到7个差异miRNA,即mmu-let-7c-5p,mmu-miR-1192,mmu-miR-294-3p,mmu-miR-29a-3p,mmu-miR-410-3p,mmu-miR-466f-3p,mmu-miR-669h-3p。结论博来霉素诱导的肺纤维化模型小鼠肺组织中存在炎性miRNA的差异表达,后者或可为肺纤维化的临床诊断与治疗提供潜在分子标志物与用药靶点。

Objective To investigate the potential effects of differentially expressed inflammatory microRNAs(miRNAs)in a mouse model of pulmonary fibrosis induced by bleomycin.Methods Twelve healthy male SPF grade C57BL/6 mice were randomly divided into control group and model group,with six mice in each group.Mice of model group were given bleomycin of 5 mg/kg by nasal inhalation,while those of control group were given an equal volune of normal saline.All mice were sacrificed fourteen days after modeling.Total RNA of lung was extracted and gene expression profiles were analyzed using mouse inflammatory miRNA PCR array plate.Target prediction and enrichment analysis were carried out for the differentially expressed miRNAs.Results Model of pulmonary fibrosis was successfully established.Seven differentially expressed miRNAs were screened out by PCR array,including mmu-let-7c-5p,mmu-miR-1192,mmu-miR-294-3p,mmu-miR-29a-3p,mmu-miR-410-3p,mmu-miR-466f-3p and mmu-miR-669h-3p.Details of target prediction and enrichment analysis are shown in results section.Conclusion This study reveals the differential expression profile of inflammatory miRNAs in a mouse model of pulmonary fibrosis.The differentially expressed miRNAs may become potential biomarkers in diagnosis,prognosis and treatment for clinical pulmonary fibrosis.