野罂粟碱对哮喘和溃疡性结肠炎模型大鼠肺及结肠组织相同差异表达基因的干预作用

Effect of intervention with Papaver nudicaule L. on the same differentially expressed genes in lung and colon tissues of asthmatic and ulcerative colitis rats

目的通过野罂粟碱调控呼吸道疾病与炎性肠病探讨中医归经理论的分子内涵,分析两类疾病动物模型在治疗后共同差异表达基因的变化从而揭示肺、肠相关联的分子机制。方法采用卵清蛋白(ovalbumin,OVA)雾化法建立过敏性哮喘模型并与复合免疫法建立的溃疡性结肠炎模型相比较,以正常大鼠作为对照。经野罂粟碱给药后,经反转录聚合酶链式反应技术(RT-PCR)检测两组模型大鼠肺组织和肠组织中共同差异表达基因mRNA表达水平。结果野罂粟碱能够明显改善大鼠肺组织和结肠组织病变,通过KEGG注释分析测序结果最终得到三个关键差异表达基因分别是C-C基序趋化因子11,碳酸酐酶1,核激素受体亚家族D组成员1,RT-PCR结果与转录组测序结果上下调情况基本一致,且野罂粟碱显著降低两组模型大鼠肺和结肠组织Ccl11 mRNA表达水平(P<0.05)及升高Ca1和Nr1d1 mRNA表达水平(P<0.05,P<0.05)。结论呼吸道疾病与炎性肠病模型大鼠的肺、肠组织具有相同差异表达基因,野罂粟能显著调节肺与大肠共同差异表达基因的水平,这可能是野罂粟归肺与大肠经的分子机制之一。

Objective To explore the molecular connotations of the TCM theory of refraction through the regulating of respiratory diseases and inflammatory bowel disease by Papaver nudicaule L., to analyze the changes of gene expression in the animal models of the two diseases after treatment, and to reveal their molecular mechanisms in the lung and intestine. Methods The allergic asthma model was established by ovalbumin(OVA) atomization and compared with the ulcerative colitis model established by the combined immunization. Normal rats were used as controls. After the administration of Papaver nudicaule L., reverse-transcription polymerase chain reaction(RT-PCR) was used to detect the expression levels of mRNA of the same differentially expressed genes in lung and intestinal tissues of the two groups of rats. Results Papaver nudicaule L. significantly improved the histological changes in the lung and colon tissues of rats. KEGG annotation analysis results for three key differentially expressed genes, C-C motif chemokine 11(Ccl11), carbonic anhydrase(Ca) 1, and nuclear receptor subfamily 1, group D, member 1(Nr1d1), and the RT-PCR results were basically consistent with the RNA-sequencing results. Papaver nudicaule L. decreased the expression of C-C motif chemokine 11 mRNA(P< 0.05) and increased carbonic anhydrase(Ca) 1 and Nr1d1 mRNA expression in the lung and colon tissues of both groups(P < 0. 05, P < 0. 05). Conclusions The lung and intestinal tissues of the rats with respiratory disease and inflammatory bowel disease have the same differentially expressed genes. Papaver nudicaule L. can significantly regulate the levels of differentially expressed genes in the lung and large intestine. This may be a molecular mechanism of Papaver nudicaule L.’s activity to locate the lung meridian and large intestine meridian.