"Golden Chamber" From The Perspective Of Melatonin, "knowing The Liver To Pass The Spleen, And Taking The Lead In Strengthening The Spleen" Mechanism

AIMAnalyzing application of diagnosis and treatment of miscellaneous diseases of“preventive treatment of disease” in Synopsis of Golden Chamber.Discussing biological mechanism of “If there is a disease of the liver,then the liver disease will be transmitted into the spleen and the spleen should be firmed first.” from the perspective of melatonin.METHODI Application analysis of “preventive treatment of disease” in Synopsis of Golden ChamberFirstly,discussing the connotation “preventive treatment of disease” based on Huang Di Nei Jing and Nan Jing by adopting the research method of philology.Secondly,analyzing the theory foundation of “preventive treatment of disease” in Synopsis of Golden Chamber based on Synopsis of Golden Chamber and Treatment of Diseases and Pulse of Zang-fu Meridians First.Finally,arranging application of “preventive treatment of disease” in the chapters of Synopsis of Golden Chamber and summarizing application regularity of “preventive treatment of disease” in Synopsis of Golden Chamber.II Mechanism research on melatonin of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” in Synopsis of Golden Chamber1.Firstly,taking “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” as an example to observe the changes of the above indices under the liver disease model based on model of rat with syndrome of liver depression,liver function biochemistry,histopathology observation,5h D-xylose excretion rate,etc.By adopting the research method of pharmacology experiment,so as to figure out the objective evidence of“If there is a disease of the liver,then the liver disease will be transmitted into the spleen.”Observing the changes of the above indices under different intervention modes by different groups and drug intervention to reflect the treatment method of “the spleen should be firmed first” to seek the objective evidence of “If there is a disease of the liver,then the liver disease will be transmitted into the spleen.”Secondly,discussing the relationship between “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” and changes of concentration of serum melatonin by testing concentration of serum melatonin.2.Exploring the intervention mechanism of melatonin on cholestatic hepatitis from metabonomics analysis and antioxidant stress mechanism based on the rat model of cholestatic hepatitis,taking melatonin as the starting point by research method of molecular biology,analytical approach of GC-MS untargeted metabolomics as well as detection technology of WES and RT-PCR,so as to expound the biological mechanism “The liver disease will be transmitted into the spleen and the spleen should be firmed first.”RESULTI Application analysis of “preventive treatment of disease” in Synopsis of Golden Chamber1.According to the analysis of original text of “preventive treatment of disease” in Huang Di Nei Jing and Nan Jing,outcomes are as follows: “preventive treatment of disease”originates from Huang Di Nei Jing and Nan Jing,and it has three meanings:(1)health preservation,and prevention before disease;(2)prevention and treatment before the disease;(3)the disease will be transmitted and cure it before change.2.According to reading of the connotation of “preventive treatment of disease” in Synopsis of Golden Chamber and Treatment of Diseases and Pulse of Zang-fu Meridians First,three traits of “preventive treatment of disease” in this chapter:(1)Taking “If one can maintain caution,he will not let evil wind go against the meridians.” as an example,it emphasizes the importance of “caution” in prevention before disease;(2)Taking guidance,expiration and inspiration,acupuncture and moxibustion and ointment massage as examples,it discusses the specific methods of curing the disease before having disease.(3)Taking sthenia-syndromes and asthenia-syndromas of liver as examples,it explains the principle of prevention of the existing disease from the two aspects of sthenia-syndromes and asthenia-syndromas to expound the disease in the liver.3.Through summarizing the application of “preventive treatment of disease” in the diagnosis and treatment of miscellaneous diseases of Synopsis of Golden Chamber,application regularities of “preventive treatment of disease” in diagnosis and treatment of miscellaneous diseases of Zhongjing:(1)Health preservation,and prevention before disease.According to the different physique of the patient and the difference of the disease,different adjustment methods such as avoiding the wind,being careful with heat,adjusting the stomach and taking medicine,etc.,should be taken respectively.The internal and external care should be taken in order to avoid the disease,such as “slight sweating and wind avoidance” after the prescription of ephedra sinica stapf,amygdalus communis vas,coix,glycyrrhiza uralensis,etc.(2)Prevention and treatment before the disease.Regardless of the severity of the disease,it is appropriate to treat it as soon as possible in order to remove the disease from the “the beginning of the disease”.For example,“He can be saved at the beginning of the disease,but he may die if there is a purulence” in pulmonray abscess of Shang Han Lun of Zhongjing.(3)The disease will be transmitted and cure it before the change.According to the deficiency of the patient's body fluid and the weakness of the splenogastric qi,it often takes care of the body fluid and supplements the middle organs at the time of the treatment of the disease.According to the truth of disease and evil,the deficiency of transmission,in order to cure the disease and not be transmitted,it should the pathways that accurately judges the disease's transmission,and first adjusts it,including magnolia officinalis and ephedra decoction,dioscorea pill,ganmai dazao decoction and so on.II Mechanism research on melatonin of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” in Synopsis of Golden Chamber1.Research on objective evidence of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” of model of rat with syndrome of liver depressionDiscussing the objective existence of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” based on the model of rats with syndrome of liver depression by different grouping intervention modes.First and foremost,“The spleen should be firmed first.” can apparently decrease the content of serum ALT,AST,ALP,TBIL and DBIL of rats with syndrome of liver depression,and remarkably improve the damage of liver tissue of rats with syndrome of liver depression as well as evidently increase 5h D-xylose excretion rate of urine of rats with syndrome of liver depression.The curative effectis better than that of simple soothing liver,simple strengthening spleen and combination of soothing liver and invigorating spleen.Thus it can be seen that “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” objectively exists.What's more,concentration of serum melatonin of rats with syndrome of liver depression can be enhanced dramatically by “firm spleen”.Firm spleen ? increase concentration of serum melatonin ? cure liver disease may become one of effective mechanism of “The spleen should be firmed first.”2.Mechanism research on “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” of melatonin? Melatonin has connection with therapeutical effect of cholestatic hepatitis,metabolism of aminoacidopathy,synthesis of glutathione.Taking melatonin as the starting point and choosing the model of cholestatic hepatitis to explore the intervention effect of melatonin on cholestatic hepatitis and the existing difference of metabolism pathways and biological mechanism of “If there is a disease of the liver,then the liver disease will be transmitted into the spleen and the spleen should be firmed first.” based on analytical approach of GC-MS untargeted metabolomics.Outcomes show that:(1)The content of cholestatic hepatitis of rat serum of ALT,AST,TBIL,DBIL,ALP and GGT can be apparently decreased by high dose of melatonin,and the change of pathology of cholestatic hepatitis of rat serum can also be dramatically improved.It has a beneficial therapeutic effect on the rat model of cholestatic hepatitis.(2)Outcomes of KEGG set pathways enrichment show that the 8 metabolic pathways are significantly different among the three groups: 1.ABC transporters;2.Glycine,serine and threonine metabolism;3.Metabolic pathways;4.Arginine and proline metabolism;5.Phenylalanine metabolism;6.Aminoacyl-tRNA biosynthesis;7.Tryptophan metabolism;8.Cysteine and methionine metabolism.Outcomes of KEGG set pathways enrichment show that the intervention effect of melatonin on cholestatic hepatitis involves metabolism pathways of amino acid,energy,carbohydrate and lipid,the degradation pathways of xenobiotics,aminoacyl-tRNA biosynthesis,the transport process of betaine and proline,glutathione and ?-beta-glucosidase.(3)Outcomes of filtering the difference of metabolism indicate that there are significant differences in 62 differential metabolites among the three groups.Combinedwith the outcomes of KEGG set pathways enrichment,seven differential metabolites related to amino acid metabolism exist in the above 8 metabolic pathways: 5-Aminovaleric acid,5-Methoxytryptamine,L-Tryptophan,threonine,glutathione,L-Methionine and indolelactate.The above seven differential metabolites are identified as potential biomarkers for melatonin intervention in cholestatic hepatitis,which is of great significance in the diagnosis of clinical cholestatic hepatitis and evaluation of the therapeutic effect of melatonin on cholestatic hepatitis.Outcomes of KEGG set pathways enrichment and filtering the difference of metabolism indicate that the therapeutic effect of melatonin on liver disease is closely related to amino acid metabolism and glutathione synthesis,providing the laboratory basis for the intervention mechanism of melatonin on liver disease.?Antioxidant stress mechanism is one of the effective mechanisms of melatonin intervention in cholestatic hepatitisTaking glutathione as the starting point and adopting the detection technology of WES and RT-PCR to explore antioxidant stress mechanism of “If there is a disease of the liver,then the liver disease will be transmitted into the spleen and the spleen should be firmed first.”Results are as follows:(1)Melatonin can significantly increase the GSH content of liver tissue of rats with cholestatic hepatitis and decrease the content of NO,MDA.The antioxidant effect is one of the important effects of melatonin on cholestatic hepatitis.(2)Melatonin can effectively raise the expression quantity of GCLM protein,GCLC mRNA and Akt mRNA of liver tissue of rats with cholestatic hepatitis,having some intervention effects on GSH synthesis.The disturbance of PI3K/Akt signal pathway is one of the antioxidant stress mechanisms of melatonin in the intervention of cholestatic hepatitis.(3)Melatonin has no significant effect on the expression of NADPH protein/mRNA in liver tissue of rats with cholestatic hepatitis,no disturbance to the dynamic of GSH reduction,and no significant effect on the expression of Nrf2.Conclusion(1)Application analysis of “preventive treatment of disease” in Synopsis of Golden Chamber“preventive treatment of disease” runs through the diagnosis and treatment of miscellaneous diseases in Synopsis of Golden Chamber.Under the guidance of “preventive treatment of disease”,Zhong Jing not only delicately prescribes in prescription,but also makes comprehensive cooperation in means of care,methods of decoction,choices of dosage,way of diet and rest and so on.Complying with the developmental trend of the disease and mastering the best treatment time of the disease to finally achieve the goal of prevention before disease,cure first,cure the disease before change with the way of according to the circumstances,no harm to its sum.(2)Mechanism research on melatonin of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.” in Synopsis of Golden Chamber(1)“If there is a disease of the liver,then the liver disease will be transmitted into the spleen and the spleen should be firmed first.” objectively exists and concentration of serum melatonin of rats with syndrome of liver depression can be enhanced dramatically by “firm spleen”.(2)Melatonin evidently improves the abnormal condition of metabolism pathways of amino acid,energy,carbohydrate and lipid of rats with cholestatic hepatitis,influencing the transport dynamics of betaine and proline,glutathione and ?-beta-glucosidase and aminoacyl-tRNA biosynthesis,especially the disturbance of amino acid metabolic pathway is the most significant.(3)Melatonin can raise the expression of glutathione synthetase,advance glutathione synthesis,activate signal pathway of PI3K/Akt and significantly improve the oxidative stress disturbance of cholestasis hepatitis.(4)Through adjusting concentration of serum melatonin,“The spleen should be firmed first.” can improve amino acid metabolism,promote glutathione synthesis,activate signal pathway of PI3K/Akt biological mechanism,so as to produce a better therapeutic effect on liver diseases.The above biological process is one of the biological mechanisms of “The liver disease will be transmitted into the spleen and the spleen should be firmed first.”...