The Application Of Metabolite Target Analysis And RT-PCR In Disease Research

Coronary artery disease (CHD) is one of the major disease which affects human health. This thesis focusing on the research of pre-diagnosis and treatment of CHD was carried out on the basis of the concept of systems biology and related technology platform, and microarray technique and bioinformatic analysis were used for screening differential expressed genes and pathways among CHD patients with different blood lipid, then the real-time RT-PCR technique was used to confirm the results of the microarray. Metabonomics were adopted for the study of CHD.Choosing CHD of different blood lipid as case study object, microarrays for gene expression profiles was futher analysised, and differentially expressed genes and pathways were got. Besides, quantitative real-time PCR (RT-PCR) was performed to verify the microarray results. ACSL1 and ADIPOQ are selected as candidate genes with satisfactory discriminatory ability. The findings suggest that ACSL1 and ADIPOQ may be of used as diagnostic indices for assessing the risk of CHD, especially for pre-diagnosis of NH-CHD.Metabolite target analysis was used for therapeutic effect evaluation. According to preliminary results of metabolomics of CHD,12 metabolites were selected from potential biomarker being discoveried in metabolomics for quantitative analysis based on normal phase HPLC-TOF/MS, The findings suggest that LysoPC(18:3), LysoPC(20:2), LysoPC(20:3), LysoPC(20:4) and DG(22:5/15:0/0:0) could be taken as potential biological markers as D-4F in treating CHD. D-4F reduced plamsa long-chain LysoPC (C≥16) may be the major reason that D-4F inhibited atherosclerosis and improved vasodialtion. Taken together, our study reveals that reducing plasma LysoPCs and DGs may be an effective approach to prevent and treat atherosclerosis.A rapid quantitative analysis method in 8 minutes of purine, pyrimidine and taurine metabolites was established based on UPLC-TOF/MS, which provides the analysis basis for target metabonomics study of diseases associated with purine, pyrimidine and taurine metabolites.