against Hepatic Encephalopathy.docx

1、against Hepatic Encephalopathyhibits Glutaminase Activity and Protects against Hepatic Encephalopathy Javier Ampuero, Isidora Ranchal, David Nuez, Maradel MarDaz-Herrero, Marta Maraver, Jos Antonio del Campo, ngela Rojas, Ins Camacho, Blanca Figueruela, Juan D. Bautista, Manuel Romero-GmezmailAbstra

2、ctAimTo investigate the influence of metformin use on liver dysfunction and hepatic encephalopathy in a retrospective cohort of diabetic cirrhotic patients. To analyze the impact of metformin on glutaminase activity and ammonia productionin vitro.MethodsEighty-two cirrhotic patients with type 2 diab

3、etes were included. Forty-one patients were classified as insulin sensitizers experienced (metformin) and 41 as controls (cirrhotic patients with type 2 diabetes mellitus without metformin treatment). Baseline analysis included: insulin, glucose, glucagon, leptin, adiponectin, TNFr2, AST, ALT. HOMA-

4、IR was calculated. Baseline HE risk was calculated according to minimal hepatic encephalopathy, oral glutamine challenge and mutations in glutaminase gene. We performed an experimental studyin vitroincluding an enzymatic activity assay where glutaminase inhibition was measured according to different

5、 metformin concentrations. In Caco2 cells, glutaminase activity inhibition was evaluated by ammonia production at 24, 48 and 72 hours after metformina treatment.ResultsHepatic encephalopathy was diagnosed during follow-up in 23.2% (19/82): 4.9% (2/41) in patients receiving metformin and 41.5% (17/41

6、) in patients without metformin treatment (logRank 9.81; p = 0.002). In multivariate analysis, metformin use H.R.11.4 (95% CI: 1.2108.8); p = 0.034, age at diagnosis H.R.1.12 (95% CI: 1.041.2); p = 0.002, female sex H.R.10.4 (95% CI: 1.571.6); p = 0.017 and HE risk H.R.21.3 (95% CI: 2.8163.4); p = 0.003 were found independently associated with hepatic encephalopathy. In the enzymatic assay, glutaminase activity inhibition reached 68% with metformin 100 mM. In Caco2 cells, metformin (20 mM) decreased glutaminase activity up to 24% at 72 hours post-treatment (p128 g/dL at 60 minute