基于RNA-Seq和生物信息学技术分析野罂粟归肺与大肠经的分子基础

‘The lung and the large intestine being interior-exteriorly related’is an important content of Chinese medicine theory,and the research between lung and large intestine channel is one of the hotspot in current studies. Previous study found that Papaver nudicaule L. had a higher enrichment in Lung and colon tissue of ulcerative colitis model rats, and caused significantly biological changes, which was indicted that the pathway of Papaver nudicaule L.has a strong relevance to the theory it belongs to the lung and large intestine channels. In this study we want to find the change regulation of massive transcription information in the lung and large intestine tissue by RNA-Seq through establishing two kinds of rat models, and extract the same differences expressing trend of genes and signal pathway.Then using bioinformatics analysis method to explore the relationship between lung and large intestine channels, reveal the molecular content of‘the lung and the large intestine being interior-exteriorly related’theory. And illuminate the essence that Papaver nudicaule L. belonging to lung and large intestine channels from the gene transcription level by the study Papaver nudicaule L. regulating the common signal pathway.

肺与大肠相表里是中医藏象学说的重要内容，研究中医肺与大肠表里经相互关系是中医理论现代化诠释的热点和难点之一。在前一个国家自然基金研究中发现野罂粟在溃疡性结肠炎模型大鼠的结肠组织和肺组织有较高富集，引起胞内的生物学变化显著，提示野罂粟的药效作用途径与其归肺与大肠经具有极强的相关性。在此基础上本课题拟通过建立肺病及肠、肠病及肺两种动物模型，全转录组测序（RNA-Seq）分析筛选两种病理变化在特定的肺组织与肠组织中海量转录信息的变化规律，提取其可能的相同差异表达趋势基因及相关信号调节通路，结合KEGG注释及通路富集分析等生物信息学方法，探寻肺与大肠经之间的实质联系，揭示肺与大肠相表里理论的分子内涵，并通过野罂粟对共同信号通路调控作用的反证，从基因转录层面阐明野罂粟归肺与大肠经的实质。

项目通过建立肺病及肠（过敏性哮喘模型）、肠病及肺（溃疡性结肠炎模型）两种动物模型，基于RNA-Seq技术探讨在病理状态下肺与大肠的相互影响与传变，结合KEGG注释及通路富集分析等生物信息学方法，探寻肺与大肠经之间的实质联系，从系统分子生物学水平部分揭示了肺与大肠相表里理论的分子内涵。实验研究发现,2273个基因在溃疡性结肠炎模型肺与大肠中有相同差异表达(p-value≤0.05,|log FC|≥1.5),其中差异基因186个基因上调,11个基因下调，20个基因在过敏性哮喘模型肺与大肠中有相同差异表达(p-value≤0.05,|log FC|≥1.5),其中差异基因13个基因上调,7个基因下调，5个基因在溃疡性结肠炎和过敏性哮喘模型肺与大肠中有共同差异表达(p-value≤0.05,|log FC|≥1.5),其中差异基因个基因上调4,1个基因下调。KEGG通路富集结果表明,差异基因参与了许多重要的生物学通路,包括ASTHMA、氮代谢、昼夜节律、趋化因子信号通路和细胞因子-细胞因子受体信号通路等。进一步的分析了3个关键的hub基因,包括REV-ERBA、CCL11、CA1这3个关键基因对于肺与大肠之间的相关信号联系可能起到了关键作用,可以作为潜在的调控位点，而他们参与的生物学信号通路可能正是肺与大肠相互关联的纽带之一。