| Polyporus umbellatus(Zhuling in Chinese)is the dried sclerotia of Polyporus umbellatus(Pers.)Fries and commonly used in clinical treatment of edema.In recent years,numerous of researches have shown the improvement of chronic kidney disease(CKD)with the extract of Polyporus umbellatus and its active components,however,the underlying mechanisms remain unclear.Renal interstitial fibrosis is the common characteristic of chronic renal failure(CRF)in the progression of various chronic kidney diseases.The development of mass spectrometry technology for identification of lipid structure has laid the foundation for the rapid development of lipidomics,a branch of metabolomics.Lipidomics is mainly performed to identify important differential lipid metabolites,and more importantly to illustrate the influence of lipid metabolism disturbance on the occurrence of diseases and reveal the improvement of lipid metabolism mechanism with drug.Here,combination of clinical data and basic research were used to investigate the disorders of lipid metabolism in CKD by using UPLC-HMDS-based lipidomics.In addition,we further explored the effect of the intervention with Polyporus umbellatus and its active component ergosterone(ERG)in chronic renal failure(CRF)rats and the mechanism on renal interstitial fibrosis by improving serum lipid metabolism disorders.Objective:UPLC-HDMS-based lipidomics was performed to explore the lipid metabolism disorder in CKD progression and prove the influence of lipid metabolism on CKD.In addition,we identify serum differential lipid metabolites in adenine-induced CRF rats,and further reveal the effects and biochemical mechanisms of PPU and ERG against CKD and tubulointerstitial fibrosis,which will provide experimental data and theoretical basis for research of Polyporus umbellatus in improvement of CKD.Methods:The present study identifies serum lipid metabolites in patients with CKD and healthy controls by using UPLC-HDMS-based lipidomics.Multivariate statistical analysis method was used to identify differential lipid metabolites in serum of patient with CKD.In addition,UPLC-HDMS-based lipidomics was performed to identify important differential lipids in adenine-induced CRF rats and analyze the correlation between lipid metabolites and creatinine levels in serum.Pharmacology,pathology,molecular biology,lipidomics and other methods were performed to investigate the underlying molecular and biochemical mechanisms of PPU and ERG intervention in CRF and against renal interstitial fibrosis.Results:1.Significant changes of lipid metabolites in patients with CKD.UPLC-HDMS-based lipidomics was performed to determine serum samples of CKD patients,additionally,chemometrics and cluster analysis methods revealed that CKD patients can be completely separated from healthy controls.We identified 77 and 49 lipid metabolites from positive and negative ion modes,respectively.Out of 126,113 identified lipid species were significantly altered in patients with CKD based on the adjusted FDR method.These results pointed to profound disturbance of fatty acid and triglyceride metabolisms in patients with CKD.The levels of free fatty acids,glycerolipids,and glycerophospholipids directly correlated with the level of serum triglyceride and inversely correlated with the levels of total cholesterol and e GFR.Logistic regression analysis showed strong correlations between serum methyl hexadecanoic acid,LPC(24:1),3-oxooctadecanoic acid,and PC(20:2/24:1)levels with e GFR and serum creatinine levels(R>0.8758).2.Disturbance of serum lipid metabolism in adenine-induced CRF rat model.We identified a total of 58 differential lipid species based on the adenine-induced CRF rat serum lipidomics,and these lipids associated with the dysregulation of arachidonic acid metabolism,sphingolipid biosynthesis,glycerophospholipid metabolism as well as bile acid biosynthesis in CRF rats.Our further study demonstrated that eight lipids including leukotrienes and bile acids presented a strong linear correlation with serum creatinine levels.3.The intervention of PPU and ERG improve renal interstitial fibrosis in CRF rats induced by adenine.Compared with control rats,the CRF rats induced by adenine showed a greatly increasing trend in serum creatinine levels and severe renal tubule-interstitial injury and renal interstitial fibrosis which illustrated that adenine-induced rats exhibited declining kidney function and tubulointerstitial fibrosis.PPU and ERG treatment caused a significantly decreased serum creatinine levels,and inhibit the secretion and deposition of extracellular matrix components.And the improvement effect of ERG is better than the improvement effect of PPU.4.The improvement of PPU and ERG in the disorder of lipid metabolites in serum of CRF.In present study,we demonstrated that the improvement of PPU and ERG in the dysregulation of eight lipid species including leukotrienes and bile acids significantly increased in adenine-induced rats,which illuminated biochemical pathogenesis of CRF and therapeutic mechanisms of PPU and ERG against renal interstitial fibrosis.Conclusion:In present study,we demonstrated the lipid metabolism disorder in the serum of CKD patients and CRF rats,and we illuminated the pathogenesis of progressive CKD is associated with disorders of lipidomic pathways including fatty acids metabolism,glycerides metabolism and glycerolipids metabolism.CRF might be associated with the disturbance of leukotriene metabolism,bile acid metabolism and lysophospholipid metabolism.The levels of leukotriene and bile acids could be used for indicating kidney function impairment in CRF.The intervention with PPU and ERG significantly ameliorated the abnormal levels of leukotrienes and bile acids.The present study reveals the biochemical mechanism of Polyporus umbellatus against renal interstitial fibrosis,which provides theoretical basis and experimental data for the rational development of the Chinese medicine Polyporus umbellatus... |
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