Acetylcholinesterase activity in an experimental rat model of Type C hepatic encephalopathy.

Patients with liver malfunction often suffer from hepatic encephalopathy, a neurological complication which can affect attention and cognition. Diverse experimental models have been used to study brain alterations that may be responsible for hepatic encephalopathy symptoms. The aim of the study was to determine whether cognitive impairment found in cirrhosis could be due to disturbance of acetylcholinesterase activity. Acetylcholinesterase activity was assessed in the brains of Wistar rats with thioacetamide-induced cirrhosis. The cirrhotic group displayed up-regulation of acetylcholinesterase levels in the entorrhinal cortex, anterodorsal and anteroventral thalamus and accumbens, whereas down-regulation was found in the CA1, CA3 and dentate gyrus of the hippocampus. Our results indicate that the experimental model of hepatic encephalopathy by chronic administration of thioacetamide presents alterations of acetylcholinesterase activity in brain limbic system regions, which play a role in attention and memory.

Portal hypertension in 18-month-old rats: memory deficits and brain metabolic activity.

Portal hypertension is a major complication of cirrhosis that frequently leads to a neuropsychiatric disorder that affects cognition. We compared the performance of 18-month-old prehepatic portal hypertensive rats (PH) and 18-month-old normal rats (CO) in spatial short-term and reference memory tasks in the Morris water maze and in active avoidance task. The PH group showed worse spatial short-term memory than the CO group. Also, the PH group tended to perform worse than the CO group in the reference memory task, but it presented a correct acquisition of the active avoidance task. We assessed the brain metabolic activity of the animals by means of cytochrome c-oxidase (COx) histochemistry. We found that the PH group developed prefrontal dysfunction characterized by increased COx activity in this region compared to the CO group. Similar results were found in the medial mammillary nucleus and dentate gyrus, whereas the CA1 area, bed nucleus of the stria terminalis, and supramammillary nucleus showed lower COx activity in the PH group as compared to the CO group. We conclude that the 18-month-old portal hypertensive rats present spatial memory impairment without alteration of implicit learning. This deficit could be related to the alteration of the metabolic activity of the brain regions involved in the processing of spatial memories.

Mammillary body alterations and spatial memory impairment in Wistar rats with thioacetamide-induced cirrhosis.

Brain tissue of patients diagnosed with hepatic encephalopathy exhibits cellular morphological changes that could be associated with memory impairment. The mammillary nuclei, located in the posterior part of the hypothalamus, are important for spatial memory formation. This work aimed to assess spatial reference memory and cellular changes in the mammillary nuclei of cirrhotic rats. Spatial reference memory of Wistar rats with thioacetamide-induced cirrhosis was assessed in the Morris water maze. Total cell number of neurons and glial cells and volume of the mammillary nuclei were quantified by stereology. Neuronal and astrocytic nuclear volume in mammillary nuclei and CA1 dorsal hippocampal subfield were assessed by nucleator probe. Cirrhotic rats showed an impaired spatial reference memory in comparison with control animals. Total number of neurons and glial cells were unaltered. In the medial mammillary nucleus (MMn), glial fibrillary acidic protein-immunoreactive astrocytes decreased in the cirrhotic group while the lateral part was unaffected. The medial part of the MMn was larger in the cirrhotic group. The cirrhotic rats showed morphometric cellular changes characterised by an increased neuronal and astrocytic nuclear volume in all the mammillary nuclei and CA1 hippocampal region. These findings suggest that cirrhotic rats show spatial memory impairment that could be linked to astrocytes and neuronal impairment in mammillary nuclei and hippocampus.

Working memory impairment and reduced hippocampal and prefrontal cortex c-Fos expression in a rat model of cirrhosis.

Hepatic encephalopathy (HE) is a frequent neurological complication observed in patients with liver malfunction. Previous studies have shown memory impairment in these patients. In order to investigate brain substrates of spatial working memory impairment in chronic HE, neuronal expression of c-Fos protein was studied in an experimental model of cirrhosis. Control and cirrhotic rats were trained on a spatial working memory task in the Morris water maze (MWM). Differences between groups were found in the working memory task. Cirrhotic rats were unable to locate the platform in the retention trial. Neuronal activation, measured by c-Fos protein, was compared between groups. No differences were found in c-Fos expression of control and cirrhotic rats that were not tested in the MWM. Working memory task produced increase in c-Fos positive cells in dorsal hippocampus, CA1 and CA3, and prefrontal cortex in control group compared to thioacetamide group or naïve, which only swam in the maze during a similar time. These findings suggest that cirrhotic rats show spatial working memory impairment that could be linked to dysfunction in neuronal activity in prefrontal cortex and hippocampus.

Diurnal locomotor activity and oxidative metabolism of the suprachiasmatic nucleus in two models of hepatic insufficiency.

Subjects with hepatic cirrhosis develop alterations of several rhythmic behavioural and biochemical patterns. Since most cirrhotic patients combine portal hypertension and hepatic impairment, our work aims to assess the extent to which rhythmical changes can be due to hepatic insufficiency or portal hypertension. This was done using two experimental models in rats, portacaval shunt model (PC) and portal hypertension by a triple stenosing ligature of the portal vein (PH). We assess diurnal locomotor activity and determine the oxidative metabolism of the suprachiasmatic nucleus (SCN) by histochemical determination of cytochrome oxidase (COX). The results show that animals with PC have altered diurnal locomotor rhythm compared to control and PH rats (p<0.001). They also present lower COX activity in the SCN (p<0.05). We conclude that rhythmic alterations are due to hepatic insufficiency and not to portal hypertension.

Cytochrome oxidase activity of the suprachiasmatic nucleus and pineal gland in rats with portacaval shunt.

Rhythmic behavioral and biochemical changes have been observed in both human and animal models with hepatic insufficiency. The basis of all these alterations is the principal endogenous pacemaker, the suprachiasmatic nucleus. The aim of this work, therefore, is to determine cytochrome c oxidase activity, a marker of neuronal activity and oxidative metabolism, in this nucleus in rats with portacaval shunt. In order to do this, this enzyme was histochemically marked and quantified by computer-assisted optical densitometry. Results show a reduced cytochrome oxidase activity in the suprachiasmatic nucleus in animals with portacaval shunts and, inversely, an increase in oxidative metabolism in the pineal gland, another circadian structure. However, the activity measured in a noncircadian brain structure, the hippocampus, which served as a control, showed no changes with surgery. Additionally, locomotor activity was assessed by actimeters and revealed a clearly reduced activity in animals with portacaval shunt. We conclude that the suprachiasmatic nucleus is possibly involved in the rhythmic changes associated with hepatic insufficiency.