| ObjectiveTo establish a reliable experimental method for the detection and analysis of peripheral blood target metabolites and metabolic profile. Preliminary evaluation of the serum level of free fatty acids (FFA) and amino acids, and the association of FFA, amino acids with severity of coronary artery lesion in different clinical types of coronary artery disease (CAD). The method of metabolic profile was used to identify metabolic markers and risk factors associated with CAD.MethodsThe serum level of FFA were determined by Gas Chromatography-Mass Spectrometry (GC/MS) combined with derivation reaction and N, O-bis (trimethylsily) trifluoroacetamide (BSTFA) was used as derivative reagent. Based upon the sample pretreatment methods and optimize the ECF derivatization reaction conditions, we established peripheral blood metabolic profile using ECF-GC/MS method for coronary heart disease (CHD) patients and healthy people.162cases were enrolled between October2010and June2011. Among them,136patients with CAD diagnosed by coronary angiography in our center and26cases of healthy volunteers as Control. CAD patients were divided into acute myocardial infarction group, unstable angina pectoris group, and stable angina group. CAD patients were underwent selective coronary artery angiography. The lesion count and Gensini score were used to evaluate severity of coronary artery. The traditional biochemical metabolites were detected. Metabolic profile analysis as well as data acquisition and preprocessing were carried out. Analysis of variance is adopted for comparison between multiple sets, Pearson is adopted for correlation analysis, and Spearman is adopted for correlation analysis when the data is non-normal distribution. Classified information and content of common peaks including FFA and amino acids were import SIMCA-P+11software for the multi-dimensional statistical analysis (the pattern recognition). PC A is adopted for pattern recognition analysis.ResultsMethodology study results showed that the established targets and metabolic profile analysis method had good precision and reproducibility and high resolution, could achieve rapid, large flux, peripheral blood metabolic targets and metabolic profile analysis.In patients with CAD, including AMI group, UAP group and SAP group, Age, gender, body mass index (Body mass index, BMI), the ratio of hypertension and diabetes, the current ratio of the number of smokers, serum TC, LDL-C, HDL-C, and taking statins and P-blockers between groups were not statistically different (P>0.05); In the AMI group, serum level of TG was lower than other groups (P<0.05); compared with the Control group, the age was older in AMI and UAP group, and the high proportion of males (P<0.01); the proportion of currently smoking in CAD patients is also higher than Control group.AMI group, UAP group and SAP group, no statistical difference in mean lesion count among the three groups (.P>0.05). There was not statistically significant difference in the proportions of one, two and three-vessel diseases among three groups (P>0.05); but with Gensini scoring, AMI group was higher than SAP group (P<0.05). Gensini scoring may be more sensitive, more comprehensively revealed the severity of coronary artery disease.linear correlation analysis was adopted for known risk factors, each component of FFAs, each component of amino acids and Gensini.The result showed, Gensini scoring were positively correlated with lesion count(r=0.553, P<0.001),age(r=0.257, P<0.001),and LDL levels(r=0.178,.P<0.05).Gensini scoring were negatively correlated with serum aspartate levels (r=-0.214, P<0.05).Compared with Contronl group, oleic acid levels increased in AMI group (P <0.05), no significant differences between groups in other FFA components (P>0.05).Compared with the UAP group, SAP group and Control group, glutamyl acid levels increased in AMI group (P<0.05) and the level of phenylalanine(P<0.01) is higher.PCA score plot of AMI group, UAP group, SAP group and Control group serum samples showed that AMI group scattered, and most overlap with Control group. SAP set of samples was significantly dispersed. The distribution of UAP group and the control group was more concentrated, but no separation trend. Four groups did not clearly separated on the score chart, indicating that no significant differences among the four groups in the metabolic spectrum.ConclusionsThe study found levels of serum aspartate were inversely related to Gensini scoring. Increased levels of serum oleic acid, glutamic acid and phenylalanine in patients with AMI. These suggesting that some components of FFAs and amino acids were related to the severity of coronary artery disease and myocardial injury. The results have certain significance for the differential diagnosis of clinical coronary heart disease, early myocardial ischemia and myocardial damage judgment. Differences metabolites or markers in CAD classification were not found by PCA analysis of serum metabolites components. ObjectiveThe mechanism of interactions of Angelica sinensis and warfarin was not clear and little attention has been paid to the effect of ferulic acid on warfarin binding with human serum albumin (HSA). The studies on supermolecular interaction of warfarin with HSA and the influence of ferulic acid on the binding would contribute to the understanding of the metabolic processes of warfarin and the effect of ferulic acid.MethodsFirstly, we analysis the content of ferulic acid in Angelica sinensis by High Performance Liquid Chromatography (HPLC) method. In order to further explore whether the interactions between warfarin and HSA occur, and the influence of ferulic acid on them, we primarily focus on investigating the effect of warfarin on fluorescence spectrum of HSA, fluorescence quenching mechanism, binding constant, Hill coefficient, binding mode, and the effect of different ferulic acid concentrations on the binding, which are expected to provide more information about the interaction between warfarin and HSA.ResultsHPLC analysis results confirmed that the average content of ferulic acid in Angelica sinensis was0.76%. This paper presents the interaction of warfarin with HSA and effect of ferulic acid on the binding by fluorescence emission and synchronous fluorescence. By decreasing binding constant and the Hill coefficient of warfarin with HSA, ferulic acid could improve the plasma concentration of free warfarin, which would increase the risk of bleeding. Under the condition of simulated human body, the free concentration of warfarin increased at least50%.ConclusionsThe results indicated that A. sinensis combined treatment with warfarin would increase the risk of bleeding. These researches help to increase understanding of the mechanism of interactions of Angelica sinensis and warfarin. And the results provide an important theoretical support for warfarin used as oral anticoagulant. |
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