| Metabolomics technology can describe the physiological and pathological changes of the body through the qualitative and quantitative analysis of small molecule metabolites.It is an effective research method to deeply understand the disease and elucidate the mechanism of drug effect.hyperlipidemia?HLP?and chronic kidney disease?CKD?both cause lipid metabolism disorders and increase the risk of a variety of cardiovascular diseases.Because of the high prevalence of these two diseases,the clinical diagnosis and treatment indicators are not sensitive enough,which often causes delays in clinical treatment.In addition,in the face of the increasing incidence of HLP and the adverse reactions of commonly used lipid-lowering drugs,how to develop safe,effective,and multi-target lipid-lowering drugs has become the primary task of preventing and treating cardiovascular and cerebrovascular diseases.As a lipid-lowering medicine,the traditional Chinese medicine Rhizoma Alismatis?RA?has been widely used in the clinical treatment of HLP by Chinese physicians,but its lipid-lowering pathway is still unclear.ObjectiveBased on the metabolomics of Ultra performance liquid chromatography-high definition mass spectrometry?UPLC-HDMS?,to find potential biomarkers of CKD and HLP respectively.This provides better indicators for the clinical diagnosis of these two diseases.Secondly,the mechanism of RA improving HLP was further studied.We hope to provide a theoretical basis and experimental basis for the rational use of RA and research and development of lipid-lowering drugs of Chinese medicine.Methods1. Potential biomarkers of CKD were searched according to metabonomics technology:16Sprague-Dawley?SD?rats were randomly divided into control?CTL?group and adenine-induced CKD?CKD?group,8 rats in each group.CKD was induced by administration of adenine(200 mg/kg·d-1)by oral gavage for 3 weeks.The control group was treated with the vehicle alone.After 3 consecutive weeks,plasma samples were collected for metabolomics research.2. Potential biomarkers of HLP were searched according to metabonomics technology:16Sprague-Dawley?SD?rats were randomly divided into CTL group and high fat diet-induced HLP?HLP?group,8 rats in each group.The CTL rats were fed with common diet during the experiment,while HLP rats were fed with high fat feed.After continuous feeding for 6 weeks,rats were then anesthetized with 10%urethane,and blood samples were obtained by carotid artery cannulation.Blood samples were used for the studies of biochemical indexes,plasma lipidomics.3.Potential biomarkers for RA intervention in HLP based on metabonomics technology:24Sixteen Sprague-Dawley?SD?rats were randomly divided into CTL group,HLP group and RA-treated HLP?RA?groups,8 rats in each group.Modeling in the same way,RA group was orally administrated with the ethanol extracts of RA(80 mg/kg/d-1),while CTL group and HLP group were orally administrated with equal volume of physiological saline.After 6weeks,blood and urine samples were taken respectively for the detection of biochemical indicators and the study of urine metabolomics.Result1.Potential biomarkers of CKD were searched according to metabonomics technology:Comparison of serum from CKD and control rats revealed significant differences in concentrations of amino acids and lipids including 33 metabolites and 35 lipid species.This was associated with marked abnormalities of fatty acid oxidation,and?-linolenic acid and linoleic acid metabolism in CKD rats.Logistic regression analysis identified tetracosanoic acid,docosatrienoic acid,PC?18:3/14:1?and L-aspartic acid,tetracosanoic acid and docosatrienoic acid as novel biomarkers of chronic interstitial nephropathy.2. Potential biomarkers of HLP were searched according to metabonomics technologyCompared with the CTL group,serum total cholesterol,triglyceride,high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were significantly abnormal in the HLP group,indicating that long-term high-fat feeding will induce HLP formation in SD rats.Based on the lipidomics of UPLC-HDMS and chemometrics methods,we identified a total of 22lipids that have abnormal changes,including 11 glycerophospholipids,6 fatty acids,2sphingolipids,1 eicosanoid,1 sterol lipid and 1 glycerolipid.These abnormal lipid metabolites indicate that in high fat diet-induced HLP rats,the metabolism of glycerophospholipids,fatty acids,sphingolipids and sterol lipids was marked perturbations.3. Potential biomarkers for RA intervention in HLP based on metabonomics technologyAbnormal serum total cholesterol,triglyceride,high-density lipoprotein cholesterol and low-density lipoprotein cholesterol in HLP group were reversed by RA.Based on the UPLC-HDMS approach and multivariate data analysis,we identified 19 metabolites as biomarkers of RA intervening in HLP.The fluctuations of these biomarkers represented the disturbances in amino acid metabolism,purine metabolism,pyrimidine metabolism and energy metabolism.After RA treatment,these perturbed metabolites were restored to normal or nearly normal levels.RA can alleviate high fat diet-induced dysfunctions in these metabolic pathways.ConclusionMetabolomics can provide more accurate and effective biomarkers than currently available methods,and help researchers to understand the pathogenesis of HLP and CKD.Metabolomics is also a powerful technical tool for in-depth study of the mechanism of traditional Chinese medicine,providing new theoretical support and experimental basis for disease diagnosis,clinical guidance of drug use,and research and development of traditional Chinese medicine. |
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