Serum Metabolic Markers And Metabolic Pathway Research On The Heart-Spleen Deficiency Syndrome Of Insomnia

Objective:Explore the serum metabolic markers and metabolic regulation pathways of heart-spleen deficiency syndrome of insomnia by using metabolomics technology,laying a foundation for the objectification research of heart-spleen deficiency syndrome of insomnia.Methods:1.Screening for insomnia patients according to the diagnostic criteria of insomnia.Screen 20 cases of insomniacs,half of whom have the most typical cardio-spleen deficiency syndrome and the rest are totally the opposite,from the research group whose patients are singled out according to the diagnostic criteria of insomnia,and 10 cases of healthy subjects were recruited in the baseline conditions of 20 cases of insomniacs.Gather the information of these subjects,such as the general condition,age,BMI index,TCM main symptoms and comorbid conditions of the patients with insomnia.2.Collect the serum of the subjects,and perform a full-spectrum detection of serum metabolomics on the heart-spleen deficiency syndrome group,non-cardiac and spleen deficiency syndrome group and the healthy group.Those data got from machine were expected to pretreatment,such as format conversion and peak extraction.The univariate analysis and multivariate analysis methods are used to screen potential metabolic markers according to the P value,VIP value and difference multiple of the comparison between groups.On the basis of the above-mentioned screening of metabolites,the same substances in the heart-spleen deficiency syndrome group/ the healthy group,the non-cardiac and spleen deficiency syndrome group/the healthy group were further identified as potential metabolic markers of insomnia;intersection objects obtained from the following three groups,namely the heart-spleen deficiency syndrome group /the health group,the noncardiac and spleen deficiency syndrome group /the health group,the heart-spleen deficiency syndrome group/the non-cardiac and spleen deficiency syndrome group were further studied,and the same substances in these three groups were identified as insomnia syndrome Class of potential metabolic markers.3.Search the metabolic markers for the human metabolic metabolite database(HMDB),determine the substance name,substance classification information,and search the Kyoto Gene and Genome Encyclopedia Database(KEGG)to determine the metabolic pathways of the differential ions.Results:1.For genders of three groups,there were 3 males and 7 females in the healthy group,2 males and 8 females in the heart and spleen deficiency syndrome group,and 3 males and 7 females in the non-cardiac and spleen deficiency syndrome group.The chi-square test X2=0.341,P=0.843,the difference between the groups was not statistically significant(P>0.05);as for the age distribution,the average age of the healthy group was 37.70±11.35,and the heart-spleen deficiency syndrome group was 33.40±13.34,and the non-cardiac and spleen deficiency syndrome group was 48.10±6.74.The age of non-cardiac and spleen deficiency syndrome group was older than the other two groups.One-way analysis of variance was F=4.866,P=0.016,and the difference between the two groups was statistically significant(P<0.05).In terms of BMI,the BMI of the healthy group was 21.54±2.44,the heart-spleen deficiency syndrome group was 20.77±3.30,the non-cardiac and spleen deficiency syndrome group was 20.04±2.11,and the one-way analysis of variance was F=0.21,P=0.81,and the difference between two groups was not statistically significant(P>0.05).2.In this study,the sleep conditions of the 10 cases of the heart-spleen deficiency syndrome group of insomnia are mainly difficult in falling asleep,easy to wake up,and not easy to fall asleep after waking up.The top 10 symptoms of the 47 most frequently occurring symptoms listed in the Insomnia TCM Symptom Survey Form are 10 times of facelessness,9 times of forgetfulness,8 times of mental fatigue,8 times of fatigue,5 times of weakness,5 times of sputum,5 times of dizziness,5 times of headache,5 times of upset,and 5 times of irritability.The sleep conditions of 10 cases of the non-cardiac and spleen deficiency syndrome group of insomnia are mainly difficult in falling asleep,dreaming,and waking up.The top 10 symptoms of the 47 most frequently occurring symptoms listed in the Insomnia TCM Symptom Survey Form are 8 times of shock,7 times of bitterness,6 times of headache,6 times of night sweats,6 times of dry mouth,6 times of fatigue,5 times of irritability,5 times of forgetfulness,5 times of facelessness,and 4 times of spontaneous sweating.3.By comparing serum metabolomics of the insomniacs with heart-spleen deficiency syndrome and the healthy subjects,we found 27 kinds of potential metabolic markers of negative ions and 258 kinds of potential metabolic markers of positive ions;By comparing serum metabolomics of the insomniacs with non-cardiac and spleen deficiency syndrome and the healthy subjects,we found 111 kinds of potential metabolic markers of negative ions and 792 kinds of potential metabolic markers of positive ions;By comparing serum metabolomics of the insomniacs with heart-spleen deficiency syndrome and the non-cardiac and spleen deficiency syndrome,we found 194 kinds of potential metabolic markers of negative ions and 1138 kinds of potential metabolic markers of positive ions.4.Single out 5 kinds of potential metabolic markers of negative ions in insomnia,78 kinds of potential metabolic markers of positive ions in insomnia,and 3 metabolic markers related to the heart and spleen deficiency syndrome in insomnia.Among them,the primary classification attribute of MG(0:0/16:0/0:0)and MG(16:0/0:0/0:0)is lipid and lipid molecules,and the secondary classification attribute is Glycerolipids,the tertiary classification attribute is monoglyceride.Glycerol 1-hexadecanoate is 1-palmitic acid monoglyceride,and the classification information of the substance has not been determined yet.The KEGG database also not showed information on the metabolic regulation pathways of the three substances.5.Among the 83 metabolic markers of insomnia,19 metabolic pathways can be found: metabolic pathway,glycerophospholipid metabolism,sphingolipid metabolism,sphingolipid signaling pathway,biosynthesis of fatty acids,biosynthesis of unsaturated fatty acids,glycine serine and threonine metabolism,D-arginine and D-ornithine metabolism,primary bile acid biosynthesis,steroid biosynthesis,glycosyl-hosphatidylinositol(GPI)anchor biosynthesis,autophagy-animal,retrograde Endogenous cannabinoid signaling,pathogenic E.coli infection,leishmaniasis,amebiasis,tuberculosis,choline metabolism in cancer,systemic lupus erythematosus.Conclusions:1.The serum metabolic markers of the heart-spleen deficiency syndrome in insomnia are 1-palmitic acid monoglyceride,MG(0:0/16:0/0:0),and MG(16:0/0:0/0: 0).2.In connection with the heart-spleen deficiency syndrome in insomnia,the main symptoms of insomnia and comorbidities may be related to abnormal metabolism of lipid molecules.Long-chain fatty acids may be the most important metabolic markers.The differences in glycerophosphate metabolism are mainly the upregulation of lecithin and cephalin.The metabolic pathway involves 19 factors such as glycerophospholipid metabolism,sphingolipid metabolism,sphingolipid signaling pathway,fatty acid biosynthesis,and unsaturated fatty acid biosynthesis and so forth.