- Email: [email protected]
Treatment of HE Through Selective Alpha 2A Adrenergic Receptor Antagonism Improves Higher Brain Function in Cirrhosis
POSTERS
Conclusions: These preliminary results suggest that the combination of partial hepatectomy and ammonia infusion is a promising new murine model of HE for investigating the mechanisms of ammonia-induced brain edema. This model would be suitable for the use of transgenic mice without presenting the problems inherent to previous murine models of HE.
CONFLICTS OF INTEREST The authors have none to declare. Corresponding author: Javier Vaquero. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.078
76
POSTER PRESENTATION
DEVELOPMENT OF A NEW ANIMAL MODEL OF OBESITY AND CHRONIC LIVER DISEASE FOR THE STUDY OF HEPATIC ENCEPHALOPATHY Rafael Ochoa-Sanchez, Mélanie Tremblay, Marc-André Clément, Marcos Ocaña-Sanchez, Christopher F. Rose Hepato-Neuro Laboratory, CRCHUM, Université de Montréal, Montréal, Canada
Background: Hepatic encephalopathy (HE) is a neuropsychiatric syndrome, a major complication of chronic liver disease (CLD/cirrhosis). With an increasing prevalence of obesity-induced cirrhosis and evidence linking bloodderived lipids to neurological impairment, we hypothesize that obesity increases the risk, severity and progression of HE. Aim: To develop and characterize an animal model of cirrhosis and obesity to investigate the synergistic effect of obesity and CLD on the development of neurological impairment and HE. M&M: Animal model of CLD and HE: The 6-week bile-duct ligation (BDL) rats, as well as sham-operated controls, were used. Obesity: To induce obesity, high-fat diet (HFD) was given for 3 weeks before BDL. Experimental groups: 1. Obese-BDL rats received HFD for 3 weeks pre-BDL and normal diet (ND) for 6 weeks post-BDL; 2. Lean-BDL rats received ND pre- and post-BDL; 3. Lean-Sham rats received ND pre- and postsham surgery. Behaviour: Motor coordination, muscular strength, and recognition memory were assessed before surgery, as well as 3 and 6 weeks post-BDL or sham surgery using the RotaRod, grip-strength and object recognition tests, respectively. Body-composition (echoMRI): Fat vs lean mass, as well as free water (ascites, fluid retention), were also monitored. S60
Results: Before the surgery, body weight and fat mass of rats on HFD (Obese-BDL) were increased in comparison to rats on ND (Lean-BDL and Lean-Sham). Three weeks after surgery, body-weight, fat mass, lean mass and free water were increased in Obese-BDL rats vs Lean-BDL rats. Long-term memory was reduced in Obese-BDL, but not in Lean-BDL, vs Lean-Sham rats. Six weeks after surgery, similar to Lean-BDL rats, Obese-BDL rats lost body weight, fat and lean mass, and increased free water vs Lean-Sham rats. Motor coordination, forelimb strength and long-term memory were impaired in Obese-BDL rats in comparison to Lean-BDL or Lean-Sham rats, whereas hind-limb strength and short-term memory were impaired in both Obese- and Lean-BDL rats when compared to Lean-Sham rats. Summary: HFD induces obesity features in healthy non-cirrhotic rats, and such effects are maintained in cirrhotic-BDL rats. Obesity also exacerbates and accelerates the accumulation of free water in cirrhotic-BDL rats. Interestingly, some neurological impairments are detected in Obese-BDL but not in Lean-BDL rats, such as long-term memory, motor coordination and forelimb strength deficits. This new animal model of CLD and obesity suggests a synergic effect, which accelerates and worsens the diseaseassociated abnormalities observed in CLD and HE. Conclusion: Obesity-induced cirrhosis in patients may result in more complex neurological manifestations, suggesting that these patients might be more susceptible to neuronal dysfunction and poor neurological performance. Therefore, this model of CLD and obesity will provide important clues to the underlying mechanisms of HE associated with obesity-induced cirrhosis and provide new insights into novel therapeutic strategies.
CONFLICTS OF INTEREST The authors have none to declare. Corresponding author: Rafael Ochoa-anchez. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.079
77 TREATMENT OF HE THROUGH SELECTIVE ALPHA 2A ADRENERGIC RECEPTOR ANTAGONISM IMPROVES HIGHER BRAIN FUNCTION IN CIRRHOSIS Helen Jones, Natalia Arias, Nathan Davies, Rajiv Jalan, Rajeshwar P. Mookerjee Division of Medicine, Institute for Liver and Digestive Health, London, United Kingdom © 2017, INASL
JOURNAL OF CLINICAL AND EXPERIMENTAL HEPATOLOGY
CONFLICTS OF INTEREST The authors have none to declare.
Corresponding author: Helen Jones. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.080
78 CIRRHOTIC PATIENTS IN ICU WITH GASTRO-INTESTINAL BLEEDING MANAGED ACCORDING TO RECENT GUIDELINES DISPLAY ALTERED BRAIN HEMOGLOBIN OXYGEN'S SATURATION ASSESSED BY NEAR INFRARED SPECTROSCOPY Maxime Mallet, Simona Tripon, Marika Rudler, Dominique Thabut, Nicolas Weiss Brain Liver Pitié-Salpêtrière (BLIPS) Study Group; Department of Neurology ICUGroupe Hospitalier Pitié-Salpêtrière-Charles Foix, Paris, France
Background and aims: Near Infrared Spectroscopy (NIRS) is a non-invasive optical technique allowing a continuous measurement of brain's hemoglobin saturation in oxygen (rSO2). It is considered as a surrogate marker of cerebral insult, and recognized as a useful tool in cardiovascular surgery and neuromonitoring. A rSO2 < 50% is associated with increased neurological impairment and post-operative mortality. In cirrhotic patients with gastrointestinal bleeding (GIB), hemoglobin (Hb) threshold for transfusion has been recently lowered to 7 g/dL. Some patients develop hepatic encephalopathy (HE) after GIB. In subarachnoid hemorrhage, a threshold of 7 g/dL of Hb could worsen neurological outcome. Objective: The aim of this study was to assess brain oxygenation using NIRS in cirrhotic patients with acute GIB admitted to ICU and managed according to recent guidelines, and to determine if brain injury was associated with Hb levels. Methods: Cirrhotic patients admitted in ICU for acute GIB were prospectively included. Bilateral continuous recording of rSO2 was started upon admission using a NIRS monitor (INVOS 5100c Cerebral Oxymeter (Covidien©) with two sensors placed on the patient's forehead. Minimal rSO2 (mini rSO2), average rSO2 (avr rSO2) and AUC of rSO2 50% (AUC50% rSO2), an integrated parameter depending on the depth/duration of desaturation under 50%, were extracted. Results: 61 patients were included (age: 60 10 years; 69%men). Etiology of cirrhosis was alcoholic 52%/viral 30%/ NASH 15%/other 3%; Child Pugh A 17%/B 28%/C 55% and MELD score 18 7. Initial Hb was 8.3 1.9 g/dL and nadir within 24 first hours was 8.0 1.8 g/dL. 33 patients (54%) had a nadir of Hb below 8 g/dL within the 24 first hours, and
Journal of Clinical and Experimental Hepatology | February 2017 | Vol. 7 | No. S1 | S22–S83
S61
POSTER PRESENTATION
Background and aims: Hepatic encephalopathy (HE) has been shown to affect daily functioning, quality of life, driving and overall mortality. However, HE is a multifactorial disease not clinically easy to assess. Many mechanisms have been proposed, one of which is altered adrenergic signaling. One animal model available for studying HE resulting from cirrhosis is chronic administration of the hepatotoxin thioacetamide (TAA) which mimics the clinical characteristics of cirrhosis including portal hypertension and HE. The aim of this study was to assess the effects of the alpha 2a adrenergic receptor (Adra2a) antagonist, BRL44408, on memory processes already altered in the TAA-induced model of cirrhosis. Methods: Rats were divided into groups: Control rats without TAA administration (CN, n = 10) and rats administered TAA intraperitoneally (IP) for 11 weeks (TAA, n = 10). On the 7th week we tested the animals (n = 10) in a reference memory task using the Barnes maze. On the 11th week, TAA animals were divided into two groups: rats treated IP with 10 mg/kg BRL44408 (BRL, n = 4) and rats treated with IP saline (SAL, n = 6). Treatments were given acutely -twice in 24 hrs with the second dose given 1hr before rats were challenged in the same task. Latencies (time to reach the hole within the Barnes maze) time spent in quadrants and locomotor activity were all studied. Results: TAA animals were not able to reach the behavioural criterion, in contrast to the CN group. No changes in locomotor activity were found between the groups. However, TAA animals treated with BRL44408 improved their latencies compared to TAA plus SAL groups. The underlying liver disease was not overtly affected by such short treatment dose as shown by a lack of significant change in liver biochemistry or haemodynamics. Conclusions: Our data suggests that selective Adra2a adrenergic receptor antagonism may be beneficial as an additional therapy for improving brain function in hepatic encephalopathy patients. Disclosures: Rajiv Jalan is a consultant to Ocera Therapeutics Inc.; has received research grants from Sequana Medical AG, Ocera Therapeutics Inc., and Yaqrit, Ltd.; has accepted speaker fees from Norgine; is involved with inventions for Ornithine Phenylacetate licensed by University College London (UCL) to Ocera Therapeutics; UCL spinout Yaqrit, Ltd., which will in-license 5 of the following inventions for which Rajiv Jalan is the main inventor (YAQ001; DIALIVE; DASIMAR: biomarker for liver failure; Neutrophil function test; TLR4 as a target for liver failure).
Conclusions: These preliminary results suggest that the combination of partial hepatectomy and ammonia infusion is a promising new murine model of HE for investigating the mechanisms of ammonia-induced brain edema. This model would be suitable for the use of transgenic mice without presenting the problems inherent to previous murine models of HE.
CONFLICTS OF INTEREST The authors have none to declare. Corresponding author: Javier Vaquero. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.078
76
POSTER PRESENTATION
DEVELOPMENT OF A NEW ANIMAL MODEL OF OBESITY AND CHRONIC LIVER DISEASE FOR THE STUDY OF HEPATIC ENCEPHALOPATHY Rafael Ochoa-Sanchez, Mélanie Tremblay, Marc-André Clément, Marcos Ocaña-Sanchez, Christopher F. Rose Hepato-Neuro Laboratory, CRCHUM, Université de Montréal, Montréal, Canada
Background: Hepatic encephalopathy (HE) is a neuropsychiatric syndrome, a major complication of chronic liver disease (CLD/cirrhosis). With an increasing prevalence of obesity-induced cirrhosis and evidence linking bloodderived lipids to neurological impairment, we hypothesize that obesity increases the risk, severity and progression of HE. Aim: To develop and characterize an animal model of cirrhosis and obesity to investigate the synergistic effect of obesity and CLD on the development of neurological impairment and HE. M&M: Animal model of CLD and HE: The 6-week bile-duct ligation (BDL) rats, as well as sham-operated controls, were used. Obesity: To induce obesity, high-fat diet (HFD) was given for 3 weeks before BDL. Experimental groups: 1. Obese-BDL rats received HFD for 3 weeks pre-BDL and normal diet (ND) for 6 weeks post-BDL; 2. Lean-BDL rats received ND pre- and post-BDL; 3. Lean-Sham rats received ND pre- and postsham surgery. Behaviour: Motor coordination, muscular strength, and recognition memory were assessed before surgery, as well as 3 and 6 weeks post-BDL or sham surgery using the RotaRod, grip-strength and object recognition tests, respectively. Body-composition (echoMRI): Fat vs lean mass, as well as free water (ascites, fluid retention), were also monitored. S60
Results: Before the surgery, body weight and fat mass of rats on HFD (Obese-BDL) were increased in comparison to rats on ND (Lean-BDL and Lean-Sham). Three weeks after surgery, body-weight, fat mass, lean mass and free water were increased in Obese-BDL rats vs Lean-BDL rats. Long-term memory was reduced in Obese-BDL, but not in Lean-BDL, vs Lean-Sham rats. Six weeks after surgery, similar to Lean-BDL rats, Obese-BDL rats lost body weight, fat and lean mass, and increased free water vs Lean-Sham rats. Motor coordination, forelimb strength and long-term memory were impaired in Obese-BDL rats in comparison to Lean-BDL or Lean-Sham rats, whereas hind-limb strength and short-term memory were impaired in both Obese- and Lean-BDL rats when compared to Lean-Sham rats. Summary: HFD induces obesity features in healthy non-cirrhotic rats, and such effects are maintained in cirrhotic-BDL rats. Obesity also exacerbates and accelerates the accumulation of free water in cirrhotic-BDL rats. Interestingly, some neurological impairments are detected in Obese-BDL but not in Lean-BDL rats, such as long-term memory, motor coordination and forelimb strength deficits. This new animal model of CLD and obesity suggests a synergic effect, which accelerates and worsens the diseaseassociated abnormalities observed in CLD and HE. Conclusion: Obesity-induced cirrhosis in patients may result in more complex neurological manifestations, suggesting that these patients might be more susceptible to neuronal dysfunction and poor neurological performance. Therefore, this model of CLD and obesity will provide important clues to the underlying mechanisms of HE associated with obesity-induced cirrhosis and provide new insights into novel therapeutic strategies.
CONFLICTS OF INTEREST The authors have none to declare. Corresponding author: Rafael Ochoa-anchez. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.079
77 TREATMENT OF HE THROUGH SELECTIVE ALPHA 2A ADRENERGIC RECEPTOR ANTAGONISM IMPROVES HIGHER BRAIN FUNCTION IN CIRRHOSIS Helen Jones, Natalia Arias, Nathan Davies, Rajiv Jalan, Rajeshwar P. Mookerjee Division of Medicine, Institute for Liver and Digestive Health, London, United Kingdom © 2017, INASL
JOURNAL OF CLINICAL AND EXPERIMENTAL HEPATOLOGY
CONFLICTS OF INTEREST The authors have none to declare.
Corresponding author: Helen Jones. E-mail: [email protected] http://dx.doi.org/10.1016/j.jceh.2017.01.080
78 CIRRHOTIC PATIENTS IN ICU WITH GASTRO-INTESTINAL BLEEDING MANAGED ACCORDING TO RECENT GUIDELINES DISPLAY ALTERED BRAIN HEMOGLOBIN OXYGEN'S SATURATION ASSESSED BY NEAR INFRARED SPECTROSCOPY Maxime Mallet, Simona Tripon, Marika Rudler, Dominique Thabut, Nicolas Weiss Brain Liver Pitié-Salpêtrière (BLIPS) Study Group; Department of Neurology ICUGroupe Hospitalier Pitié-Salpêtrière-Charles Foix, Paris, France
Background and aims: Near Infrared Spectroscopy (NIRS) is a non-invasive optical technique allowing a continuous measurement of brain's hemoglobin saturation in oxygen (rSO2). It is considered as a surrogate marker of cerebral insult, and recognized as a useful tool in cardiovascular surgery and neuromonitoring. A rSO2 < 50% is associated with increased neurological impairment and post-operative mortality. In cirrhotic patients with gastrointestinal bleeding (GIB), hemoglobin (Hb) threshold for transfusion has been recently lowered to 7 g/dL. Some patients develop hepatic encephalopathy (HE) after GIB. In subarachnoid hemorrhage, a threshold of 7 g/dL of Hb could worsen neurological outcome. Objective: The aim of this study was to assess brain oxygenation using NIRS in cirrhotic patients with acute GIB admitted to ICU and managed according to recent guidelines, and to determine if brain injury was associated with Hb levels. Methods: Cirrhotic patients admitted in ICU for acute GIB were prospectively included. Bilateral continuous recording of rSO2 was started upon admission using a NIRS monitor (INVOS 5100c Cerebral Oxymeter (Covidien©) with two sensors placed on the patient's forehead. Minimal rSO2 (mini rSO2), average rSO2 (avr rSO2) and AUC of rSO2 50% (AUC50% rSO2), an integrated parameter depending on the depth/duration of desaturation under 50%, were extracted. Results: 61 patients were included (age: 60 10 years; 69%men). Etiology of cirrhosis was alcoholic 52%/viral 30%/ NASH 15%/other 3%; Child Pugh A 17%/B 28%/C 55% and MELD score 18 7. Initial Hb was 8.3 1.9 g/dL and nadir within 24 first hours was 8.0 1.8 g/dL. 33 patients (54%) had a nadir of Hb below 8 g/dL within the 24 first hours, and
Journal of Clinical and Experimental Hepatology | February 2017 | Vol. 7 | No. S1 | S22–S83
S61
POSTER PRESENTATION
Background and aims: Hepatic encephalopathy (HE) has been shown to affect daily functioning, quality of life, driving and overall mortality. However, HE is a multifactorial disease not clinically easy to assess. Many mechanisms have been proposed, one of which is altered adrenergic signaling. One animal model available for studying HE resulting from cirrhosis is chronic administration of the hepatotoxin thioacetamide (TAA) which mimics the clinical characteristics of cirrhosis including portal hypertension and HE. The aim of this study was to assess the effects of the alpha 2a adrenergic receptor (Adra2a) antagonist, BRL44408, on memory processes already altered in the TAA-induced model of cirrhosis. Methods: Rats were divided into groups: Control rats without TAA administration (CN, n = 10) and rats administered TAA intraperitoneally (IP) for 11 weeks (TAA, n = 10). On the 7th week we tested the animals (n = 10) in a reference memory task using the Barnes maze. On the 11th week, TAA animals were divided into two groups: rats treated IP with 10 mg/kg BRL44408 (BRL, n = 4) and rats treated with IP saline (SAL, n = 6). Treatments were given acutely -twice in 24 hrs with the second dose given 1hr before rats were challenged in the same task. Latencies (time to reach the hole within the Barnes maze) time spent in quadrants and locomotor activity were all studied. Results: TAA animals were not able to reach the behavioural criterion, in contrast to the CN group. No changes in locomotor activity were found between the groups. However, TAA animals treated with BRL44408 improved their latencies compared to TAA plus SAL groups. The underlying liver disease was not overtly affected by such short treatment dose as shown by a lack of significant change in liver biochemistry or haemodynamics. Conclusions: Our data suggests that selective Adra2a adrenergic receptor antagonism may be beneficial as an additional therapy for improving brain function in hepatic encephalopathy patients. Disclosures: Rajiv Jalan is a consultant to Ocera Therapeutics Inc.; has received research grants from Sequana Medical AG, Ocera Therapeutics Inc., and Yaqrit, Ltd.; has accepted speaker fees from Norgine; is involved with inventions for Ornithine Phenylacetate licensed by University College London (UCL) to Ocera Therapeutics; UCL spinout Yaqrit, Ltd., which will in-license 5 of the following inventions for which Rajiv Jalan is the main inventor (YAQ001; DIALIVE; DASIMAR: biomarker for liver failure; Neutrophil function test; TLR4 as a target for liver failure).