| Chinese medicine processing is a unique traditional pharmaceutical technology in China,and has been listed as one of the national intangible cultural heritage.As we all know,Chinese medicine is mostly used in the form of compound prescriptions,but the current research on Chinese medicine processing is mostly conducted on single flavor tablets,and this research mode is disconnected from the characteristics of Chinese medicine clinical compound prescriptions,which have many flavors and complex relationships,and it is difficult to clarify the mechanism of them when researching.Therefore,the study of the mechanism of the processing of Chinese herbal medicines in pairs not only takes into account the combination factors,but also avoids the difficulty of elucidating the mutual influence of multiple drugs due to their complex relationships,and is more in line with clinical practice.Dried ginger and schisandra are classical pairs commonly used in clinical treatment of cough and asthma,and their medicinal properties can be moderated through processing,and the concoction is mostly used in clinical medicine,but there are cases of mixed use of raw and prepared products for cough and asthma.Therefore,the study of the efficacy changes and mechanism of action before and after the processing of dried gingerschisandra pair is crucial for the correct and rational application of clinical concoctions,and is also a scientific issue worth exploring.Objective: The method of gut microbiota and bile acid metabolism,combined with the investigation of pharmacodynamic indexes,was used to investigate the mechanism of action of the concoction-combination,in order to clarify the scientific connotation of the concoction-combination and provide reference for clinical treatment and other studies.Method:1 Differences in the pharmacological efficacy of dried ginger and schisandra before and after processing in rats with treating cold asthma The SD male rats were randomly divided into normal group,model group,GW group(dried ginger-Schisandra group),PC group(roasted ginger-vinegar Schisandra group)and dexamethasone group,12 rats in each group.The rat model of cold asthma was established by the method of "OVA + ice water bath".The histopathological changes of the lungs in each group were investigated by HE method;the eosinophils and neutrophils in the alveolar lavage fluid were counted;the contents of Ig E,IL-13,IL-4,TNF-α and INF-γ in the serum and alveolar lavage fluid of each group were measured,and the differences in the pharmacological effects of the dried ginger and schisandra in rats with cold asthma were investigated before and after the processing.2 Differences in gut microbiota of dried ginger and schisandra before and after processing in rats with treating cold asthma The 16 S r DNA sequencing was used to sequence the rat feces of each group,and combined with bioinformatics analysis and statistical methods,the species composition of each group and the species diversity between groups were analyzed to screen the differential groups at the genus level and compare the differences in the degree of backregulation of the differential groups before and after the concoction of dried gingerschisandra.3 Differences in bile acid metabolism of dried ginger and schisandra before and after processing in rats with treating cold asthma The bile acid content in the feces of each group of rats was detected by LC-MS/MS,combined with chemical pattern recognition and statistical methods to screen out the differential bile acids,compare the changes of differential bile acid content in the feces before and after the concoction of dried ginger-schisandra chinensis,and perform correlation analysis on the differential flora and differential bile acids by Spearman method to investigate the regulatory effect of gut microbiota on bile acids.Results: 1 Differences in the pharmacological efficacy of dried ginger and schisandra before and after processing in rats with treating cold asthma The lung tissues of rats in the model group showed increased thickness of bronchial walls,narrowing of lumen and irregular shape,etc.After administration,the walls became thinner and the mucus secretion in the lumen was reduced,and the improvement effect of the medicine to raw group was stronger than that of the medicine to concoction group;the eosinophil and neutrophil counts of the model group were significantly higher than those of the normal group,and all showed a tendency to be retuned after administration,and the retuning tendency of dried ginger-schisandra was lower than that of the medicine to raw group.The ELISA results showed that the concentrations of Ig E,IL-13,IL-4 and TNF-α in the model group were significantly higher than those in the normal group,and the concentrations of INF-γ were significantly lower than those in the normal group,and all of them showed a tendency to be retuned after administration,and the retuning tendency in the medicine to raw product group was stronger than that in the medicine to concoction group.2 Differences in gut microbiota of dried ginger and Schisandra before and after processing in rats with treating cold asthma The 16 S r DNA analysis of gut microbiota in cold asthmatic rats showed that the alpha diversity indices chao1,observed,PD whole tree and Shannon were significantly lower in the model group compared with the control group;compared with the model group,the alpha diversity indices of mice in each administration group were significantly increased.Ten differential bacterial groups were screened out,among which the relative abundance of Allobaculum,Candidatus_Arthromitus,Dubosiella,Lactobacillus,Ligilactobacillus,Romboutsia,and Rothia was significantly reduced in the model rats compared with the normal group.The relative abundance of Alloprevotella,EscherichiaShigella,Streptococcus in the model rats was significantly increased compared with the normal group;all of them were retraced to different degrees toward the normal group after administration,while the retracing trend was less obvious in the case of baked ginger-vinegar Schisandra than in the case of dried ginger-Schisandra.3 Differences in bile acid metabolism of dried ginger and Schisandra before and after processing in rats with treating cold asthma By OPLS-DA analysis,it was found that the model group was clearly separated from the normal group with the farthest distance,and the distance between the medicine pair raw product group and the normal group was the most similar,followed by the dexamethasone group and the medicine pair concoction group;statistical analysis of screening five differential bile acids revealed that the content of DCA was significantly down-regulated in cold-drinking volvulus asthmatic rats,and the content of ACA,GLCA,GCDCA,GUDCA was significantly The levels of ACA,GLCA,GCDCA and GUDCA were significantly down-regulated,and the levels of ACA,GLCA,GCDCA and GUDCA were significantly up-regulated.Spearman correlation analysis showed that upregulation of Allobaculum,Candidatus_Arthromitus,and Ligilactobacillus increased DCA content and decreased GUDCA,ACA,GLCA,and GCDCA content;upregulation of Alloprevotella Dubosiella,Lactobacillus,Romboutsia,Streptococcus,Escherichia-Shigella may cause a decrease in DCA content and an increase in GUDCA,ACA,GLCA,GCDCA;an increase in Rothia may cause a decrease in DCA,GUDCA content decreased and the content of ACA,GLCA,GCDCA increased.Conclusion: Dried ginger-schisandra chinensis before and after processing improved the physiological and inflammatory indexes,increased the richness and diversity of gut microbiota,and regressed the differential flora to the normal group;interfered with the abnormal bile acid metabolism to the normal group in rats with cold asthma.The ameliorative effect of roasted ginger-vinegar Schisandra on treating cold asthma rats was weaker than that of dried ginger-Schisandra. |
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