Serum Metabonomics Study Of Nervous Tinnitus Patients With Kidney Deficiency Syndrome

Objectives:Using super high performance liquid chromatography and series-level 4 pole flight time mass spectrometry (UPLC-Q-TOF/MS) metabolomic research technology combined with Ingenuity pathways analysis (IPA) to study the metabolic spectrum features and molecular network biological pathways in nervous tinnitus patients with kidney deficiency syndrome upon those different metabolites, and attempt to explore the molecular mechanisms of nervous tinnitus kidney deficiency syndrome.Methods:The subject mainly adopt a novel design that the cross-section study to explore and a further longitudinal study to prove the mechanisms of nervous tinnitus kidney deficiency syndrome. Firstly, we find out the differential metabolites related to nervous tinnitus by UPLC-Q-TOF/MS and then construct some molecular network biological pathways based on those differential metabolites with IPA in the cross-section study. Secondly, some differential metabolites related to nervous tinnitus and molecular network biological pathways were obtained from the longitudinal study applying same research ways. Lastly, comparing with the datas from both clinical trials, we got some common differential metabolites and molecular network biological pathways which were more relative with nervous tinnitus.The details as follows:serum samples of 50 nervous tinnitus kidney deficiency syndrome and 50 healthy volunteers were collected in the cross-section study. All of the 22 participants from the longitudinal study were same ones and we gathered serum samples of them in the states of before and after tinnitus. After that, both principal components analysis (PCA) and orthogonal partial least squares-discrimination analysis(OPLS-DA) were used to tidy data. Finally, some potential metabolic biomarkers related to nervous tinnitus kidney deficiency syndrome were found upon metabolic analysis, and many networks as well as canonical pathways were built according to all of the differential metabolites by IPA to further understand the pathogenesis of nervous tinnitus.Results:1. There was no significant difference of age, sex, BMI, blood pressure, blood glucose, blood fat, hepatic and renal function and so on between nervous tinnitus and the healthy in both cross-section study and longitudinal study.2. There were 12 differential metabolites identified in nervous tinnitus patients in the cross-section study. Up regulated ones were glutamic acid,5-HT, choline, betaine, arginine, orotic acid,8-oxoguanine, phenylethylamine, LysoPC (18:0), and down regulated ones were inosine, taurine, octanoic acid. Above identified metabolites might be relative to some changes, such as amino acid metabolism, small molecule biochemistry, cell signaling, cellular compromise, infectious disease, neurological disease, energy production, lipid metabolism, molecular transport. Applying molecular biology network platform for differential metabolites analysis, and the results showed the probable biology canonical pathways were choline degradation Ⅰ, calcium signaling, phospholipases, uridine-5’-phosphate biosynthesis, pyrimidine ribonucleotides De Novo biosynthesis, neuropathic pain signaling in dorsal horn neurons, alanine degradation Ⅲ, alanine biosynthesis Ⅱ,4-hydroxyphenylpyruvate Biosynthesis, aspartate biosynthesis, superpathway of methionine degradation, nNOS signaling in neurons, proline degradation, glutamate biosynthesis Ⅱ, glutamate degradation X.3. In the longitudinal study, we find out 10 differential metabolites in nervous tinnitus group. The up regulated ones were glutamic acid,5-HT, sarcosine, orotic acid,8-oxoguanine, and the down regulated ones were taurine, L-asparagine, aconitic acid, malonaldehyde, Glycyl-L-leucine. Above identified metabolites might be relative to some changes, such as molecular transport, small molecule biochemistry, nervous system development and function. Applying molecular biology network platform for differential metabolites analysis, and the results showed the probable biology canonical pathways were asparagine biosynthesis Ⅰ, calcium signaling, uridine-5’-phosphate biosynthesis, pyrimidine ribonucleotides De Novo biosynthesis, circadian rhythm signaling, neuropathic pain signaling in dorsal horn neurons, tRNA charging, Huntington’s Disease signaling, taurine biosynthesis, methylglyoxal degradation Ⅰ, GABA receptor signaling, L-cysteine Degradation Ⅲ, L-cysteine degradation Ⅰ,4-aminobutyrate Degradation Ⅰ, glutamate receptor signaling.4. Comparing the results from both cross-section study and longitudinal study, there were 5 common differential metabolites found in nervous tinnitus kidney deficiency syndrome, which were glutamic acid,5-HT, taurine,8-oxoguanine, orotic acid. Similarly, the common biology canonical pathways were calcium signaling, pyrimidine ribonucleotides De Novo biosynthesis, uridine-5’-phosphate biosynthesis, neuropathic pain signaling in dorsal horn neurons.Conclusion:1. Differential metabolites were obtained between nervous tinnitus and healthy volunteers based on UPLC-Q-TOF/MS, PCA scores plots had the trends to discriminate these two groups.2. As the common differential metabolites found in nervous tinnitus kidney deficiency syndrome among cross-section study and longitudinal study, Glutamic acid,5-HT, taurine,8-oxoguanine and orotic acid, having the same change in disease, were possible the potential metabolic biomarkers which could play a predictive indication in the incidence of nervous tinnitus.3. Molecular network biological analysis platform was able to effectively study differential metabolites of nervous tinnitus kidney deficiency syndrome. Calcium signaling, pyrimidine ribonucleotides De Novo biosynthesis, uridine-5’-phosphate biosynthesis and neuropathic pain signaling in dorsal horn neurons were common biology canonical pathways found in both cross-section study and longitudinal study, and perhaps that they had an important influence on nervous tinnitus.4. The method of one cross-sectional study to explore the pathogenesis of disease together with another verification of the longitudinal research, might be more effective to know the information related to disease, which is worth further discussion and reference.