Differential Metabolites In Cirrhotic Patients With Hepatitis B And Muscle Mass Loss

Background: Sarcopenia is a progressive and diffuse loss of muscle mass,muscle strength,and function.Pre-sarcopenia is defined as low muscle mass without loss of muscle strength and poor physical performance.It leads to complications(infections,hepatic encephalopathy,ascites)and poor overall survival in cirrhosis patients.The metabolic disorder may be the root cause of low muscle mass in cirrhotic patients.However,the metabolites and mechanism of hepatitis B virus(HBV)-related liver cirrhosis with pre-sarcopenia remain unclear.This study aimed to reveal the metabolic profile and identify potential biomarkers of muscle mass loss in hepatitis B virus(HBV)-related decompensated liver cirrhosis and provide directions for early nutritional intervention.Method: Patients with HBV-related liver cirrhosis and healthy control people were recruited between August 2021 and June 2022 at the Affiliated hospital of Qingdao University.The skeletal muscle mass index(SMI: cm~2/m~2)was equal to the skeletal muscle area(SMA: cm~2)divided by height squared(m~2).Muscle mass loss was defined as an SMI less than 46.96 cm~2/m~2 for males and less than 32.46 cm~2/m~2 for females.Morning fasting blood was collected and centrifuged at 3000 × g for 15 min.The plasma was stored at-80°C until analyze.Liquid chromatography-mass spectrometry(LC-MS)and gas chromatography-mass spectrometry(GC-MS)analyses were conducted to find differential metabolites.The unsupervised principal component analysis(PCA),partial least-squares-discriminant analysis(PLS-DA),and orthogonal partial least-squares discriminant analysis(OPLS-DA)were used to identify the stability,repeatability,and difference between groups.Differential metabolites and pathways were detected with variable importance on projection(VIP)scores > 1 and P values < 0.05 based on the KEGG database.Receive operator characteristic(ROC)curves were used to further identify the key metabolites.Results:1.GC-MS single metabolic platform: 20 HBV-related decompensated liver cirrhosis patients with muscle mass loss were designated Group S,20 HBV-related decompensated liver cirrhosis patients with normal muscle mass were designated Group NS,and 20 healthy people were designated Group H.Twenty-five pathways were significantly different in the plasma of Group S compared with Group NS.Strong predictive value of 11 metabolites(inosine-5’-monophosphate,phosphoglycolic acid,D-fructose-6-phosphate,N-acetylglutamate,pyrophosphate,trehalose-6-phosphate,fumaric acid,citrulline,creatinine,(r)-3-hydroxybutyric acid,and 2-ketobutyric acid)were selected as potential biomarkers in Group S patients compared with Group NS patients.Two pathways may be associated with loss of muscle mass in patients with liver cirrhosis: amino acid metabolism and central carbon metabolism in cancer.2.LC-MS single metabolic platform: 20 HBV-related decompensated liver cirrhosis patients with muscle mass loss were designated Group S and 20 HBV-related decompensated liver cirrhosis patients with normal muscle mass were designated Group NS.Three hundred and twenty-five differential metabolites were detected,only 32 of them were imported into 5 disturbed pathways.These disturbed pathways were associated with choline metabolism,primary bile acid biosynthesis,phenylalanine metabolism and glycerolipid metabolism.After we adjusted the P value,there were 30 differential metabolites between Group S and Group NS.L-Phenylalanine and rhodexin A were down-regulated in Group S,and other metabolites were up-regulated.3.LC-MS ad GC-MS dual metabolic platform: 15 HBV-related liver cirrhosis patients with pre-sarcopenia were designated as Group S;Fourteen liver cirrhosis without pre-sarcopenia were designated as Group NS.Five pathways and twenty-eight pathways were defined as disturbed pathways in the plasma of liver cirrhosis patients with pre-sarcopenia by LC-MS and GC-MS,respectively.Most of these pathways are related to amino acid metabolism.Forty-two differential metabolites were imported into the disturbed pathways.Moreover,3-hydroxypropanal,hydrocinnamic acid,betaine aldehyde,phosphohydroxypyruvic acid,(r)-3-hydroxybutyric acid,and creatinine were identified as potential biomarkers for HBV-related liver cirrhosis with muscle mass loss.Conclusion: Certain biomarkers were distinguished between muscle mass loss and normal muscle mass in HBV-related cirrhosis patients.The pathways of amino acid metabolism,central carbon metabolism in cancer,urea cycle,and glyoxylate and dicarboxylate metabolism may be associated with muscle mass loss in patients with HBV-related liver cirrhosis.Differential metabolites detected in this study provide a direction for nutritional supplementation in liver cirrhosis.