| OBJECTIVE:In recent years,studies have found that the intestinal and respiratory tract have different physiological structures and functions,but under pathological conditions,especially Asthma patients often Coexist with Inflammatory bowel increase(IBD).Although some patients do not have a history of acute or chronic respiratory disease,many patients with IBD have impaired lung function,and more and more studies have found that respiratory diseases are closely related to the occurrence of inflammatory bowel disease.Papaver nudicaule L.has been used clinically for cough and diarrhea in various ethnic minorities in the north.In the previous study,the main active ingredient of wild papaverine,antitussive and antidiarrheal,was found to have higher concentration in the colon tissue and lung tissue of model rats,which may further explore the possibility of mutual transmission of respiratory diseases and intestinal diseases.The mechanism of action and the relationship between Papaver nudicaule L.control and the two diseases.This study combines the latest research progress and previous research results of modern medicine on the interaction between respiratory and intestinal diseases,and further adopts transcriptome sequencing technology(RNA-Sequencing,RNA-Seq)combined bioinformatics analysis to screen the same differentially expressed genes and enrichment pathways in lung and intestinal tissues and the regulation of Papaver nudicaule L.in the "lung disease and intestine" model and the "gut disease and lung" modelMETHODS:The allergic asthma model was replicated by OVA atomization and the lung disease and intestinal and "gut disease and lung" models were established in comparison with the composite immunological replication ulcerative colitis model.Normal-rats were used as controls.60 SPF SD rats were 200～220 g male and female,aged 8-10 weeks,randomly divided into five groups:blank control group(C),ulcerative colitis model group(UCM),ulcerative colitis administered group(UCT),the asthma model group(ASM),and the asthma administered group(AST),of which 12 were in the normal control group,and 12 in each group.The three treatment groups of UCT and AST were intragastrically administered daily,and the C,UCM,and ASM groups were intragastrically administered with equal doses of sterile water,and the disease activity index(DAI)of the rat was observed daily.After successful modeling,the rats in the model group were sacrificed.The same position of each colon was selected for HE staining to observe the inflammation of the lungs and colon of each group.RNA was extracted from rat lung tissue and intestinal tissue at the same time.RNA-Seq technology was used to screen out differentially expressed genes and signaling pathways,and reverse transcriptase polymerase chain reaction(RT-PCR)was used to detect the administration of wild papaverine.Changes in mRNA expression levels of differentially expressed genes in lung tissue and intestinal tissues of the latter two groups of rats.RESULTS:The study found that after the treatment of the model,the activity of the rats was reduced,the hair color was gray and messy,and the rats in the asthma model group showed a significant increase in respiratory stimuli,especially during nebulization.In rats with ulcerative colitis,the number of stools and stools without stools increased.In this study,the KEGG annotation analysis results in the final three key differentially expressed genes:CC motif chemokine 11(CCL11),Carbonic Anhydrase(CA)1,nuclear receptor subfamily 1,group D,member 1(Nr1d1),the results of RT-PCR were consistent with the up-regulation of transcriptome sequencing results,and it was found that wild papaverine can significantly improve lung and colon tissue lesions in rats.Wild papaverine significantly decreased the expression of CCL11 mRNA(P<0.05)and increased CA1 and Nr1d1 mRNA expression in lung and colon tissues of the two groups(P<0.05,P<0.05).Conclusion:This study established a model rat and used RNA-Seq technology to analyze the transcriptome data of lung tissue and colon tissue.The results of the study were the molecular mechanisms of the transformation of lung disease and intestinal and intestinal disease and lung.The exploration of poppy regulation and the relationship between the classics provides a certain foundation.This study has the following conclusions:(1)The lung disease model and the enteropathy model were replicated separately.After the administration of Papaver nudicaule L.,the pathological manifestations of the two disease model rats showed a recovery trend,indicating that Papaver nudicaule L.had certain therapeutic effects on asthmatic rats and ulcerative rats.(2)Constructed a rat model of allergic asthma and a rat model of ulcerative colitis,and identified the intestinal expression in the rat model of ulcerative colitis.The differentially expressed genes in the blank control group compared with the ulcerative colitis model were 2459.In the lung tissue,there were 362 differential genes in the blank control group compared with the ulcerative colitis model;186 genes in the lung tissue and intestinal tissue were differentially expressed,of which 175 were up-regulated and 11 were down-regulated.In the allergic asthma model,compared with the blank control group,there were 210 differentially expressed genes in the intestinal tissue;163 differentially expressed genes in the lung tissue;20 differentially expressed genes in the lung tissue and intestinal tissue,among which 13 genes were up-regulated and 7 genes were down-regulated.Three common differentially expressed genes were screened by comparing the differentially expressed genes of the two groups,CC motif chemokine 11 CC motif chemokine 11(CCL11),carbonic anhydrase 1 Carbonic Anhydrase(CA)1,nuclear hormone receptor Member of group D of the subfamily 1 nuclear receptor subfamily 1,group D,member 1(Nr1d1).(3)According to the GO annotation and KEGG pathway annotation analysis,it is preliminarily clarified that the intestinal and respiratory systems are linked by signaling pathways such as nitrogen metabolism,circadian rhythm,asthma pathway,immune system,and cytokine-cytokine receptor interaction.(4)The common differentially expressed genes based on the two sets of models,combined with RT-PCR results,showed that the CCL11 gene was up-regulated;in addition,the CA1 and Nr1d1 genes were significantly down-regulated,which was consistent with the RNA-Seq results.Up-regulation of genes involves processes such as chemokine pathways.Down-regulation of genes involves regulation of homeostasis and regulation of circadian rhythm.It is proved that CCL11,CA1 and Nr1d1 genes and their signaling pathways may be the molecular mechanism of the "lung and large intestine phase" theory.First,the regulation of CCL11,CA1 and Nr1d1 genes by Papaver nudicaule L.may also be one of the molecular basis for exerting antitussive and diarrhea and returning to the lungs and large intestine. |
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