| Objectives: This study aims to explain the component targets and mechanism of action of Danggui Shengjiang Yangrou Decoction(DSYD)in the intervention of IBS-D due to spleen-kidney yang deficiency,to predict the possible categories of its scientific connotation using network pharmacology and molecular docking,and to search for relevant scientific evidence using methods such as metabolomics and gut microbiota abundance combined with physiological and biochemical indicators detection.Methods:1.In this study,the network pharmacology method combined with ultra-high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UHPLC-LTQ Orbitrap MS)was used to screen and analyze the relationship between drug ingredients,targets,pathways and diseases.The network was visualized,and the molecular docking technology was used to verify the affinity and binding mode of its core components and potential core targets,and the key targets and pathways of DSYD acting on spleen-kidney yang deficiency syndrome models were excavated.2.Taking DSYD as the test drug,the clinical manifestations,physiological and biochemical changes,and organ pathological changes of experimental rats with spleenkidney yang deficiency and irritable bowel syndrome fed with DSYD for 30 days were observed.Combined with the BDI-GS evaluation system,the effect of Danggui Ginger Mutton Soup on experimental rats was evaluated.3.Taking female adult SD rats as experimental objects,a rat model of spleen-kidney yang deficiency was established,and the paws,tails,and tongues of the model and experimental rats were analyzed by Image J image analysis software,and the clinical changes of spleen-kidney yang deficiency syndrome was studied in combination with open field experiments.4.16 s rDNA high-throughput sequencing technology was used to obtain intestinal microbial diversity data for analysis,and RT-PCR was used to detect the changes of energy metabolism and inflammation related gene expression in brown adipose tissue,such as UCP,DIO,S100 b,AMPK,etc.To explore the therapeutic mechanism of DSYD in improving spleen-kidney yang deficiency rats with irritable bowel syndrome.Results:1.Network pharmacology and molecular docking confirmed the reliability of the core components of DSYD acting on potential core targets of IBSDSYD acted on AKT1,INS,TNF,TP53,IL6,EGFR,CTNBB1,and IL1 B through 8core components,including turmeric ketone,calamone,palmitic acid,urophyllic acid,β-Myrrhene β-Sitosterol,stigmasterol,oleic acid;inhibited pro-inflammatory factors through AGE-RAGE signaling pathway,c AMP signaling pathway,MAPK signaling pathway,IL-17 signaling pathway,and Ca M signaling pathway;alleviated the inflammatory response,enhanced the intestinal mucosal barrier and regulated intestinal motility,and played a role in improving the disease targets related to spleen-kidney yang deficiency.2.DSYD corrected the physiological and biochemical indicators of IBS-D model rats with spleen and kidney yang deficiencyCompared with the normal group,the white blood cells and platelets of model rats were significantly increased,with a significant difference(P <0.05).After oral administration of DSYD,there was no significant difference in white blood cells and platelets compared to the normal group.It can be seen that DSYD can restore white blood cells and platelets to normal.The bilirubin and triglycerides of model rats were significantly reduced,and the urea nitrogen and creatinine levels were significantly higher than those of the normal group(P <0.05).Combined with the results of urine routine tests,the urine of the spleen-kidney yang deficiency group showed significant acidification changes.After taking DSYD,the urine p H value,urine protein,and biochemical indicators tended to normal.The BDI values of the liver,spleen,lungs,kidneys,brain,ovaries,and adrenal glands of female rats fed with DSYD for 30 days were all>1,indicating that the administration of DSYD helped to restore the coefficients of various organs,while the BDI values of the thymus in female rats were all<1,indicating that adenine and senna leaf water caused inhibition of thymus function in female rats,and DSYD had no significant improvement effect.However,the total cumulative GS values of all organs in female rats were>10.3.Administration of DSYD improved the clinical symptoms of IBS-D model rats with spleen and kidney yang deficiencyCompared with model rats,the mental state,ear and tail color,hair,and feces of the low,medium,and high dose groups of DSYD improved significantly(P <0.05).Through open field experiments,it was found that the low,medium,and high dose groups of DSYD showed significant improvements in time,distance,rapid movement time,and slow movement time in the central and surrounding areas compared to the model group.From the RGB value data,DSYD can improve the color of the tail,sole,and tongue of ISB-D rats.The q PCR results showed that compared with the normal group,the expression levels of IL-6,IL-1β,and TNF-α related to acute enteritis were significantly increased(P <0.05),while the expression of IFN-γ was significantly decreased(P <0.05).,the expressions of IL-6,IL-1β,and TNF-α tended to be normal after being fed with DSYD which were significantly different from the model control group(P <0.05),and the expression of IFN-γ was significantly increased.4.DSYD regulated the abundance and structure of gut microbiota in IBS-D model rats with spleen-kidney yang deficiency,and alleviated inflammatory symptoms.By regulating serum c AMP and T3 levels,energy metabolism was influenced through pathways such as AGE-RAGE,MAPK,IL-17,and Ca M signaling pathwaysThe experiment observed that the temperature and activity of the model rats decreased,the temperature recovered and the activity increased after feeding DSYD,and the serum c AMP and T3 levels increased with the increase of the dosage,which played an important role in thermoregulation;In terms of gut microbiota abundance,the spleenkidney yang deficiency group showed a significant decrease in the Firmicutes phylum,which is a group of bacteria that can produce high amounts of short chain fatty acids.Some members of the Firmicutes phylum can help digest cellulose and other difficult to digest polysaccharides,thereby providing energy.Therefore,the anal temperature of the spleen-kidney yang deficiency model animals decreased.However,after feeding DSYD the bacterial population that produces high levels of short chain fatty acids increased to81.6%-84.6%,resulting in a return to normal body temperature;Different gut microbiota have different effects on the regulation of the host immune system,and this interaction may play an important role in maintaining intestinal immune balance and preventing the occurrence and development of inflammatory bowel disease.By improving the structure of gut microbiota,the symptoms of inflammatory bowel disease can be effectively alleviated.On the other hand,compared with the normal group,the energy metabolism related gene calsyntenin3 β,DIO2,S100 B,and leptin in IBS-D model rats with spleen-kidney yang deficiency syndrome were significantly down-regulated(P <0.05),and the three genes were significantly up-regulated after oral administration of DSYD,with significant differences compared to model rats(P <0.05).The expression level of uncoupling protein in the model group was significantly higher than that in the normal group,but three genes were significantly up-regulated after administration of DSYD,and there was significant difference compared with the model rats(P <0.05),but there was no significant difference compared with the normal group,which further affected the level of energy metabolism and increased body temperature.Conclusion: The results of network pharmacology research suggest that DSYD can enhance the intestinal mucosal barrier and regulate intestinal motility through pathways such as AGE-RAGE signaling,c AMP signaling,MAPK signaling,IL-17 signaling,and Ca M signaling,thereby improving the target of IBS in spleen-kidney yang deficiency syndrome.The IBS-D model of spleen-kidney yang deficiency can be successfully modeled by intragastric administration of adenine and senna leaf water.After oral administration of DSYD,it can correct the physiological and biochemical indicators of model rats and improve their clinical manifestations.After studying the gut microbiota and energy metabolism,it was found that the model group showed a significant decrease in Firmicutes and Bacteroidetes,leading to a decrease in anal temperature in the model animals.However,after feeding DSYD,the microbiota that produced high levels of short chain fatty acids recovered and increased to 81.6%-84.6%,resulting in a return to normal body temperature.At the same time,it was found that DSYD significantly affected the energy metabolism related gene calsyntenin3 β,DIO2 and S100 B and further promoted the recovery of body temperature.By improving the structure of the gut microbiota,it can affect the expression of inflammatory factor genes and restore intestinal health. |
PDF Full Text Request