南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (1): 39-46.doi: 10.12122/j.issn.1673-4254.2021.01.05

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博莱霉素诱导小鼠肺纤维化中差异表达的 mRNA 及其与LncRNA的共表达网络关系

喻雪飞,李 理,郑林鑫,李伟峰   

  • 出版日期:2021-01-26 发布日期:2021-01-26

Differential mRNA expression in C57BL/6 mice with bleomycin-induced pulmonary fibrosis and its association with LncRNA co-expression network

  • Online:2021-01-26 Published:2021-01-26

摘要:

目的 研究博莱霉素诱导的小鼠肺纤维化模型中mRNA/LncRNA表达谱的变化,筛选出肺纤维化相关的mRNA,与差异LncRNA进行编码-非编码共表达(CNC)生物信息学分析。方法 采用气管内注射博来霉素诱导的小鼠肺纤维化模型。SPF级C57BL/6小鼠随机分为模型组(10只/组,向心端气管内注入100 μL博来霉素溶液3 mg/kg)及对照组(10只/组,注入等量的0.9%的氯化钠注射液),于14 d时处死大鼠留取肺组织。通过Masson染色和HE染色评估组织肺纤维化程度。利用LncRNA芯片技术筛选肺纤维化过程中差异表达的mRNA与LncRNA表达谱,通过NCBI数据库、UCSC数据库等对差异mRNA进行GO及pathway 等生物信息学分析,筛选出纤维化可能相关的 mRNA,通过 qRT-PCR 验证其表达变化,进一步与所有差异表达LncRNA进行CNC共表达分析,构建纤维化可能相关mRNA与差异LncRNA共表达网络。结果 模型组肺组织纤维化程度显著高于对照组。基因芯片显示与对照组相比,模型组中表达上调的mRNA有 127个,表达下调的有184个,GO及pathway分析发现差异表达的基因在生物学功能上主要涉及免疫反应、细胞分化、细胞骨架等;参与的信号通路主要有细胞因子与细胞因子受体相互作用、趋化因子信号转导通路等。生物信息学分析发现纤维化相关mRNA与差异表达的LncRNA存在显著的共表达网络关系。结论 本研究筛选出小鼠肺纤维化发生过程中差异变化mRNA/LncRNA表达谱,生物信息学分析得出纤维化相关mRNA确实与大量差异的LncRNA存在高度相关的共表达关系。

关键词:

Abstract:

Objective To study the changes in mRNA and long non-coding RNA (lncRNA) expression profiles in a mouse model of bleomycin-induced lung fibrosis and identify lung fibrosis-related mRNA for coding-noncoding coexpression (CNC) bioinformatics analysis of the differential lncRNAs. Methods Lung fibrosis was induced by intratracheal injection of bleomycin in 10 C57BL/6 mice and another 10 mice with intratracheal injection of saline served as the control group. Lung tissues were harvested from the mice at 14 days after the injections and lung fibrosis was assessed using Masson and HE staining. LncRNA chip technology was used to screen the differentially expressed mRNAs and lncRNAs in mice with lung fibrosis, and GO and KEGG pathway analyses of the differential mRNAs were performed using NCBI database and UCSC database to identify possible fibrosis-related mRNAs, which were validated by qRT-PCR to construct a coding and non-coding co- expression network with the differential lncRNAs. Results Compared with the control mice, the mice with intratracheal injection of bleomycin showed obvious lung fibrosis. The results of gene chip analysis showed that 127 mRNAs were up-regulated and 184 mRNAs were down-regulated in the model group as compared with the control group. GO and pathway
analysis suggested that the differentially expressed genes participated mainly in immune response, cell differentiation, and cytoskeletons; the involved signal pathways were associated mainly with cytokine and cytokine receptor interaction and chemokine signal transduction. Bioinformatics analysis identified a significant coexpression network between the fibrosis-related mRNA and the differentially expressed lncRNA. Conclusions In mice with lung fibrosis, the differential expressions of fibrosis-related mRNAs in the lung tissues are closely correlated with the co- expressions of a large number of differential lncRNAs, which points to a new direction for investigation of the pathogenesis of pulmonary fibrosis.

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