| Chronic bronchitis (CB), is a disease that cause serious inflammation of trachea, bronchial mucosa and other lung tissue, accompanied with long term cough and short breath. It is import to study the mechanism of chronic bronchitis, identify related potential biomarkers and apply them to diagnose this disease.We used different concentrations of tobacco smoke to model chronic bronchitis rats, in order to find better experiment condition. Based on LC-MS and NMR metabolomics, sera were analyzed to identify the potential biomarkers and their metabolic pathways. Dexamethasone and bergenin were administrated to treat chronic bronchitis and we applied advanced technologies to explore the mechanism on the disease. The results are as follow:In order to find better condition to model chronic bronchitis, we set 3 concentrations of smoke, that is 40%,60% and 80%, and monitored the process of modeling using HOPE 8050 container. According to the result, there were remarkable infiltrations of macrophages and neutrophils in peribronchial, perivascular and alveolar of TS rats compared with the rats of the control group, particularly in the 60% TS group. In addition, hemosiderin was detected in the lung tissues of rats in the 60% TS group, but not in other groups. Therefore,60 %TS was considered as the most suitable condition to establish CB models within 4 weeks.To study the mechanism of CB induced by tobacco smoke, LC-MS was applied to analyze the changes of metabolites in serum. Through the advanced technologies combined with multivariate analysis such as PCA and PLS-DA,11 metabolites were identified including Among them, the levels of lysophosphatidylethanolamine (18:1), lysophosphatidic acid (18:1), lysophosphatidylethanolamine (18:0), lysophosphatidylethanolamine (16:0), lysophosphatidylethanolamine (20:4), docosahexaenoic acid,5-hydroxyindoleacetic acid and 5’-carboxy-γ-tocopherol were higher in TS group compared to control group. Conversely, the levels of 4-imidazolone-5-propionic acid,12-hydroxyeicosatetraenoic acid and uridine were lower in TS group (Student’s t test, p< 0.05). The results indicated that the mechanism of CB was related to amino acid metabolism and lipid metabolism, particularly lipid metabolism. In addition, lysophosphatidylethanolamines were proved to be important mediators, which could be used as biomarkers to diagnose CB.To better understand the mechanism of CB, NMR technology was used to find more potential biomarkers. Dexamethasone and bergenin were administrated to treat the disease and NMR metabolomics was applied to find the dynamic changes in metabolites while dosing the medicine. The metabolic profiles were analyzed by multivariate statistical analysis, including PCA, PLS-DA and OPLS-DA models. The PCA score showed a good separation between control and model groups (R2X= 0.965 and Q2= 0.858). The PLS-DA score showed that control group and model group were separated along t[1] (R2X= 0.318, R2Y= 0.98, Q2= 0.87). Based on the VIP, correlation coefficient and p value, a variety of metabolites were identified between control and CB group, including serine, alanine, glutamine, proline, glutamate, lactate, creatine phosphate, glucose, isoleucine, leucine, valine, acetone, acetoacetate, pyruvate, glycine, threonine, isobutyrate and glycerol. The dynamic fluctuations of those biomarkers in sera from different groups were detected. The results also suggested that the anti-inflammatory mechanism of dexamethasone may be associated with glycine and threonine metabolism and glycolysis while bergenin could change BCAA metabolism including leucine, isoleucine and valine, glycine, serine and threonine metabolism, and glycolysis to treat CB. |
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