The Establishment Of Heyi Rat Model And The Blood Metabolomics Of Study

Objective:Based on Mongolian medicine theory,to explore the method of establishing the pathological model of Heyi and to find its related detection indexes through animal experiments.Through the study of blood metabolomics in model animals and Heyi constitution,the mechanism of Heyi was discussed.Methods:1.Based on the four pathogenic factors in Mongolian medicine theory,SD rats were divided into normal group,low,medium and high dose model group,and each group was divided into male group and female group,8 groups in total.The normal group was given a normal diet.The model group was given different doses of buckwheat,coffee and black tea every day for 4 months.In the fourth month,a certain amount of wind,disturbance,shock and alternate hot and cold environment were applied.The general state,food intake and body weight of the rats were observed.To calculate the viscera index;the contents of phosphocreatine kinase isoenzyme(CK-MB),phosphocreatine kinase(CK)and lactate dehydrogenase(LDH)were determined by biochemical analyzer.ELISA was used to detect 5-HT,y-aminobutyric acid(GABA),norepinephrine(NE)and dopamine(DA)in the brain.HE staining was used to observe the histopathological changes of heart tissues,and immunohistochemistry was used to determine transforming growth factor TGF ?-1(TGF-?-1)and endothelin 1(ET-1)in heart tissues.Differential metabolites and their distribution trends in blood were analyzed by orthogonal partial least squares discriminant analysis(OPLS-DA)using liquid-mass spectrometry.2.In the study of blood metabolomics of Heyi constitution patients,according to the Mongolian medicine theory and the constitution judgment of relevant experts,30 Heyi constitution patients and 30 polymeric constitution patients were selected from the volunteers participating in the project,and the metabolic differences in the two groups of human blood were detected and compared by using the liquid-mass combination technology,and their metabolic pathways were analyzed.Results:I.Compared with the normal group,the model rats in each dose group gradually changed significantly in many aspects during the 4-month period.In terms of general status and food intake,the rats' hair was rough and dull,their activities were flexible,they were short-tempered and easily frightened,their stools were dark and shiny,and their food intake decreased.In terms of body weight,compared with the normal group,there was a slight reduction in the low-dose group of male rats,and a significant reduction in both the medium-dose and high-dose groups.The female rats in the low,medium and high dose groups had only slight weight loss.In terms of organ index,compared with the normal group,the organ index of some model groups(kidney in the low-dose male group,lung in the low-dose female group,liver and spleen in the medium-dose female group)decreased.In terms of biochemical indexes,compared with the normal group,the cardiac tissues of the medium-dose group CK-MB,CK,the male high-dose group CK-MB,CK and LDH were significantly improved.In the female medium group CK,and high dose group Ck-MB,CK,and LDH were significantly increased.The levels of 5-HT,GABA,NE and DA in the brain tissues of rats in each dose model group were significantly increased.In terms of pathological observation,the myocardial fibers were closely arranged and orderly in the normal group,while in the high-dose model group,a small amount of myocardial fiber nuclei were seen to contract,myocardial fiber fractures and a small amount of bleeding.Immunohistochemical results showed that the expression trend of TGF-?1 and ET-1 was low dose>medium dose>high dose>normal group.The results of serum metabolomics in rats showed that,compared with the normal group,29,33 and 43 differential metabolites and 10,12 and 16 related metabolic pathways were found in the male model group with low,medium and high doses,respectively.In the female low-dose,medium-dose and high-dose model groups,36,32 and 31 differential metabolites and 12,11 and 14 related metabolic pathways were found,respectively.Compared with the normal group,57 differential metabolites and 18 related metabolic pathways were found in the male group.In the female group,59 differential metabolites and 17 related metabolic pathways were found.In both sexes,compared with the normal group,a total of 75 differential metabolites and 21 related metabolic pathways were found in the model group.2.The results of serum metabolomics analysis of Heyi constitutions showed that compared with the polymeric constitutions,a total of 19 differential metabolites were identified and 20 related metabolic pathways were found,among which 3 metabolic pathways were significantly different,namely Pyrimidine metabolism,Nitrogen metabolism and Sphingolipid metabolism pathways.Conclusion:1.The model of Heyi can be established by giving rats a certain amount of soba,coffee and black tea and a certain amount of wind,disturbance,shock and alternate cold and hot environment.2.The general status,food intake,body weight,cardiac CK-MB,CK,LDH and brain 5-HT,GABA,NE and DA of the animals could be selected as the detection indicators.3.The 75 differential metabolites and 21 metabolic pathways were found in the blood metabolites of Heyi mouse model and the blood metabolites of Heyi constitution can be used as the basic data for further screening of the human markers and pathogenesis of Heyi.4.The results of this study laid an experimental foundation for the pathogenesis research of Heyi disease,provided detection indexes for the research on the mechanism of action of Heyi Mongolian medicine,and provided research ideas and specific methods for the research on the characteristic pathological model of Mongolian medicine.