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TDK conference 2018Balajthy Bálint - year 6 University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry Supervisors: Dr. Zoltán Bárány, Dr. Dávid Sándor Kiss Hepatic encephalopathy is a neuropsyhiatric syndrome caused by portosystemic shunt and/or liver failure. The pathogenesis of the disease has not yet been accurately detailed, though it is well known that cerebral pathogenesis is linked with astrocytes. Certain aspects of the pathogenesis of HE has previously been examined at our department in vitro, using primary cell cultures where the capacity of proinflammatory cytokine production by astrocytes was measured with the application of agents proved to take part in the pathogenesis of HE. The results were not consistent and required further investigations. In this study, the previously mentioned in vitro model was adopted to measure the extracellular concentration of tumor necrosis factor (TNF)-α by ELISA. For one of the astroglia cell cultures, brain tissues from Sprague-Dawley rats aged 1-2 days were isolated, then prepared with cytosine β-d-arabinofuranoside (Ara-C) and L-leucine-methylester (LME) to reduce the number of microglia cells. In addition, a new model was set up to by shaking the astrocyte cultures in order to reduce the number of microglia. To assess the effectiveness of shaking, immunocytochemical techniques were used to measure the decrease in number of microglia. Investigations were carried out to determine the effect on the production of TNF-α of hydrogen peroxide, ammonia, manganese and bacterial lipopolisaccharides as causative agents of oxidative stress in pure rat primary astrocyte cultures. The results made necessary the examination of the cell viability using the above mentioned agents. Our results have shown that both of the cell cultures were able to produce TNF-α in physiological conditions, however TNF-α levels were reduced by the application of the examined factors. In addition, the reduction in the production of TNF-α was not obviously explained by the results of cell viability examinations. List of lectures |