Research On The Biological Mechanism Of Brain-gut Dialogue In "Seeing Liver Disease, Knowing The Liver To Pass The Spleen, And Taking The Lead In Rehabilitating The Spleen"

AIMDiscussing the objective evidence of“knowing the liver disease will be transmitted into the spleen,strengthening the spleen first" in Synopsis of Golden Chamber.Exploring the "knowing the liver disease will be transmitted into the spleen "based on the new mechanism of intestinal chromaffin cell brain-gut dialogue The biological mechanism of "spreading the spleen,taking the spleen first".METHODThis study used pharmacological research methods,chronic restraint and subcutaneous injection of carbon tetrachloride(CCL4)to establish a liver depression and chronic liver injury model.The Sini San group used Sini San to intervene for 42 consecutive days,representing the treatment principle of "treating the liver".The Liujunzi Decoction group used Liujunzi Decoction to intervene for 42 consecutive days,representing the treatment principle of strengthening the spleen".The Chaishao Liujunzi Decoction A group used Chaishao Liujunzi Decoction to intervene for 42 consecutive days,representing the treatment principle of " treating the liver and strengthening the spleen ".The Chaishao Liujunzi Decoction B group used Chaishao Sini San to intervene in the first 21 days,and Chaishao Liujunzi Decoction to intervene in another 21 days,representing the treatment principle of " treating the liver first".The Chaishao Liujunzi Decoction C group used Chaishao Liujunzi Decoction to intervene in the first 21 days,and Chaishao Liujunzi Decoction to intervene in another 21 days,representing the treatment principle of "strengthening the spleen first".The model group,the normal group and the positive drug group were given intragastric administration of distilled water or drugs of the same volume.This study observed the general state of rats,using light microscopy to observe the pathology of rat liver tissues,and detected serum liver function biochemistry serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),lactate dehydrogenase(LDH),total bilirubin(TBIL),direct bilirubin(DBIL),Total protein(TP),albumin(ALB),which can evaluate the rat liver function,ELISA method was used to detect rat serum diamine oxidase(DAO),gastrin(GAS)content to observe the rat gastrointestinal motility function and intestinal mucosal barrier function.By observing the changes of the above indicators in different intervention methods,we seek objective evidence of "knowing the liver disease will be transmitted into the spleen,strengthening the spleen first".From the perspective of brain-gut relationship,ELISA method was used to detect the content of substance P(SP)in rat serum and colon,and RT-PCR method was used to detect the expression of Ghrelin and VIP mRNA in rat hypothalamus and colon tissue to observe each group.Changes in brain-gut peptide expression were observed by immunohistochemical staining in the number and morphology of colonic intestinal chromaffin cells(EC cells).Based on the brain-gut dialogue mechanism,we discuss some of the biological mechanisms of "knowing the liver disease will be transmitted into the spleen,strengthening the spleen first" in Synopsis of Golden Chamber.RESULT1.From the analysis results of the general state of rats in each group,on the basis of pathological manifestations of liver depression and the liverliver cannot stretch,rats with liver depression and chronic liver injury also experienced burnout,lethargy,depression of strong thread,symptoms of spleen deficiency syndrome such as loose stool;the biochemical manifestation of serum liver function in model rats is elevated in ALT,AST,ALP,DBIL,TBIL,LDH,and decreased in ALB and TP,indicating that the liver function of rats is damaged.The decrease of gastrointestinal hormone GAS and the increase of DAO indicate that the gastrointestinal motility function and intestinal mucosal barrier function of model rats are impaired,confirming the objective existence of "seeing liver disease,knowing liver disease will be transmitted into the spleen ".2.The Chaishao Liujunzi Decoction C group used the treatment plan of "strengthening the spleen first,then treating the liver and strengthening the spleen",which represented the treatment principle of " strengthening the spleen first ".The "strengthening the spleen first " treatment method significantly improved the abnormal mood and behavior of rats.From the perspective of liver histopathology and biochemistry of serum liver function in rats,the " strengthening the spleen first " treatment reduced the levels of ALT,AST,ALP,DBIL,TBIL,LDH,increased the levels of ALB and TP,and improved the liver function.and liver damage status.From the analysis of the results of gastrointestinal hormones,the " strengthening the spleen first "treatment increased GAS content and reduced DAO content,and promoted the recovery of gastrointestinal motor function and intestinal mucosal barrier function in rats.The curative effect of the Chaishao Liujunzi Decoction C group is better than that of the other administration groups.It is confirmed that " knowing the liver disease will be transmitted into the spleen,strengthening the spleen first " objectively exists.3." knowing the liver disease will be transmitted into the spleen,strengthening the spleen first " is related to the regulation of brain-gut peptides.Using brain-gut peptide as the entry point,the content of substance P(SP)in rat serum and colon was detected by ELISA method,and the expression of Ghrelin and VIP mRNA in rat hypothalamus and colon tissue was detected by RT-PCR method.The results showed that the relative expression levels of Ghrelin mRNA in the colon and hypothalamus tissues of the model group decreased,the relative expression levels of VIP mRNA increased,and the SP colonic expression in the rat colon increased while the serum SP content decreased.The Chaishao Liujunzi Decoction C group represented the " strengthening the spleen first " can improve the abnormal expression of Ghrelin,VIP,SP of cerebral intestinal peptide,indicating that the Chaishao Liujunzi Decoction C group represents the intervention effect of"strengthening the spleen first " on liver depression combined with chronic liver injury is related to the regulation of brain-gut peptide expression.4."knowing the liver disease will be transmitted into the spleen,strengthening the spleen first" is related to the brain-gut dialogue mechanism of intestinal enterochromaffin cells.Immunohistochemical staining was used to observe the number and morphological changes of colonic intestinal enterochromaffin cells(EC cells).The results showed that the number of 5-HT immunoreactive intestinal enterochromaffin cells in the colon tissue of the model group increased and the morphology was abnormal.The "strengthening the spleen first " represented by the Chaishao Liujunzi Decoction C group reduced the number of intestinal enterochromaffin cells,improved the morphology of intestinal enterochromaffin cells,and helped to restore the brain-gut dialogue mechanism of intestinal enterochromaffin cells.The regulation of the brain-gut dialogue mechanism of intestinal enterochromaffin cells by the Chaishao Liujunzi Decoction C group representative treatment is one of its intervention mechanisms for liver stagnation and chronic liver injury.CONCLUSION1.When the model of liver depression combined with chronic liver injury rat liver function is damaged,gastrointestinal motor function and intestinal mucosal barrier function are also affected," knowing the liver disease will be transmitted into the spleen " is objective.2.The treatment methods of " Treating the liver,Strengthening the spleen,Treating the liver and Strengthening the spleen,Treating the liver first,strengthening the spleen first " can improve the liver function and gastrointestinal motility function of liver depression and chronic liver injury model rats to varying degrees.The treatment of "strengthening the spleen first"has the best curative effect." knowing the liver disease will be transmitted into the spleen,strengthening the spleen first " objectively exists.3.The pathogenesis of " knowing the liver disease will be transmitted into the spleen " is related to abnormal regulation of Ghrelin,VIP and SP.4.The treatment of " knowing the liver disease will be transmitted into the spleen,strengthening the spleen first " can regulate the brain-gut axis mechanism by regulating the levels of Ghrelin,VIP,SP of brain-gut peptides in the rat model of liver depression and chronic liver injury.5.The treatment of " knowing the liver disease will be transmitted into the spleen,strengthening the spleen first " can regulate the number of intestinal enterochromaffin cells,regulate its synthesis and release of 5-HT function,interfere with the neurotransmitter conduction and feedback of intestinal enterochromaffin cells.The intervention effect of " knowing the liver disease will be transmitted into the spleen,strengthening the spleen first" is closely related to the brain-gut dialogue mechanism of intestinal enterochromaffin cells.