Research On Serum Metabolic Profiling Of Cirrhosis Patients With Minimal Hepatic Encephalopathy And The Intervention Effect Of Rhubarb Decoction

Objective: Using GC-TOFMS metabonomics and UPLC-QTOFMS analysis platform in combination with bioinformatics and pattern recognition analysis method,to analyze the sera metabolite profilings of all selected subjects to find out the related biomarkers;to reveal the mechanism of MHE cognitive dysfunction and the association and the relationships and evolution of metabolites in the development of related diseases,to explore the effect of rhubarb decoction on serum metabolic profile of MHE patients.Methods: In this study,a total of 107 subjects serum samples were collected,include: 50 healthy volunteers(NC group),12 decompensated cirrhosis patients(CS group),12 decompensated cirrhosis patients(DS group),29 HBV-induced cirrhosis patients with minimal hepatic encephalopathy(MHE group),4 hepatic encephalopathy patients(HE group),and the 29 patients with MHE were randomly divided into 15 groups as a traditional Chinese medicine group and the other 14 patients as a placebo group,and then using rhubarb decoction and sterile water for injection to retain enema respectively,Once a day,treated for 7 days,collected the serum before and after treatment.Using the detection method of GC-TOFMS and UPLC-QTOFMS technology,the subjects’ serum samples were tested for metabolomics,combined with the Principal Component Analysis(PCA)and Orthogonal Partial Least Squares-Discriminant Analysis(OPLS-DA)to analyze the pattern,and then using the variance analysis,t test and other statistical methods for significant differences in metabolites test.Results: Compared with NC group,the CS、DS、MHE and HE groups were gradually separated.There was no differential metabolites were found in NC group and CS group,66 differential metabolites were decreased between NC group and the DS group,60 differential metabolites were decreased between NC group and the MHE group,73 differential metabolites were decreased between NC group and the HE group,compared with the NC group,there were 27 common differential metabolites in the DS,MHE and HE groups,the increased of these differential metabolites including adenine,phenylalanine and so on;and the decreased metabolites including sphingosine,vitamin E and so on.Compared with the NC group,the DS and MHE group differential metabolites were generally close in number,but the MHE group of various bile acids and bile salts and other metabolites content about 2 times higher than the DS group,while the contents of branched chain amino acids and sphingosine and other metabolites were lower than those of DS group.MHE and HE group metabolites in the number and concentration levels were significantly different,the number of metabolites in HE group was 13 more than that in MHE group,serum levels of carnitine、nerve acid and other metabolites in the serum of the HE group were higher than those in the MHE group,the levels of valine,sphingosine and other metabolites were significantly lower than those of MHE group.There was no significant difference in the metabolite profile of MHE patients before and after treatment with rhubarb decoction and sterile water for injection.Conclusion:(1)Compared with NC group,the CS、DS、MHE and HE groups with the gradual progress of the disease,the corresponding significant differences in the corresponding metabolites;(2)There was no differential metabolites were found in NC group and CS group;(3)The patients of the MHE and DS group differential metabolites were generally close in number,while the levels of bile acids and bile salts in the MHE group were higher than those in the DS group;the MHE patients cognitive function changed may related to BACC/AAA ratio imbalance and sphingosine content reduction and other factors;(4)the number and concentration of metabolites in HE group were significantly higher than those in MHE group,so identifying the MHE patients early and taking intervention measures timely have important practical significance for the prevention of MHE into CHE;(5)There was no significant difference in the metabolite profile of MHE patients before and after treatment with rhubarb decoction and sterile water for injection.