CC BY
949th multidisciplinary international
Conference of Biological Psychiatry
«Stress and Behavior»
Proceedings of the 9th International Multidisciplinary Conference «Stress and behavior» Saint-Petersburg, Russia, 16-19 May 2005 Editor: Allan V. Kalueff, PhD
CONFERENCE ABSTRACTS 5. PSYCHONEUROIMMUNOLOGY
APOPTOSIS IN LYMPHOCYTES OF ALCOHOLIC PATIENTS
O.Yu. Fedorenko, A.N. Bokchan, N.M. Rakitina, E.G. Karzanova, S.A. Ivanova Mental Health Research Institute, Tomsk, Russia Programmed cell death and its morphologic manifestation of apoptosis is essential mechanism in adult organisms to maintain normal cellular homeostasis and reject defective cells. Although the occurrence of apoptosis has been known for decades, it is only recently that ethanol has been found to trigger widespread apoptosis (Olney et al., 2001). Alcoholism is associated with variety of clinical issues related to both mental health and physical health effects. Prolonged use of ethanol results in dependence, and discontinuation of ethanol produces a severe withdrawal syndrome (SWS) marked by anxiety, ataxia, hyperalgesia, seizures, coma and even death (Jung et al., 2005). Ethanol-induced tissue injury and cell death have been extensively studied from the periphery to the central nervous system. Ethanol can damage virtually every organ and tissue because of its ability to alter membrane integrity and affect key biochemical processes throughout the cells. Human monocytes isolated from healthy subjects after binge alcohol drinking, as well as the brain tissue of alcoholics, showed evidence of apoptotic neuronal cell death. (Freund 1994; Singhal et al., 1999). In connection with the above-stated the objective of this study was researching the apoptosis of lymphocytes in alcoholic patients with SWS and after being treated. Methods 24 men 31—57 years old with alcoholism (average flow of disease was 15 years) were examined along with 20 healthy age-matched men. The inclusion criterion was alcoholism (diagnosed according to ICD10: F10 — alcoholism). The examination of alcoholics was carried out in dynamics: at receiving patients with SWS (first point) and after 2-week treatment (second point). Lymphocytes were isolated from fresh heparinized venous blood of alcoholic patients and healthy controls, as adopted method with gradient ficoll-gipak. The content of cells expressing the Fas-receptor as apoptosis marker was studied by indirect immunofluorescent method with use of monoclonal antibodies to CD95-antigen and the results were expressed in percentage terms. Also determination of apoptosis by morphological examination was performed. For that blood smears from heparinized venous blood onto a glass slide were prepared. The slides were air-dried for 30 min prior to staining and fixing with May-Grunwald and Giemsa solutions. A minimum of 200 cells were examined for morphological changes characteristic of apoptosis (nuclear condensation, vacuolation, and blebbing), with the use of light microscopy. The results were expressed in percentage terms.
Results and discussions: The relative content of bloodstream lymphocytes expressing receptors of readiness to Fas-dependent apoptosis (CD95) in alcoholics with SWS and after their treatment (16.32 ± 1.39% and 15.43 ± 5.12% respectively) significantly exceeds the same index in healthy controls (12,00 ± .77%, p < 0.05). The absolute content of CD95 lymphocytes in alcoholic patients before and after treatment (555.44 ± 106.38 x 106 cells/L and 373.35 ± 82.43 x 106 cells/L) was also higher as compared with healthy individuals (123.48 ± 20.43 x 106 cells/L, p < 0.05). Together with the readiness to apoptosis, assessment of lymphocytes with morphological signs of apoptosis was conducted. The percentage and the absolute content of lymphocytes with morphological characteristics of nuclear fragmentation in alcoholic patients was twice as much in comparison with the corresponding indexes of healthy men. The percentage content of apoptotic lymphocytes in alcoholic patients was 2.68 ± 0.46% before treatment vs 2.15 ± 1.93% after (0.97 ± 0.35% in controls, p <0.05). The absolute content of apoptotic lymphocytes in
Psychopharmacol. Biol. Narcol. 2005. Vol. 5, N 2. P. 910
Psyhopharmacology & biological narcology
ISSN 1606-8181
alcoholic patients was 53.78 ± 10.07 x 106 before and 63.37 ± 16.14 x 106 cells/L after treatment (9.98 ± 0.65 x 106 cells/L in controls, p < 0.05). Also the index of apoptosis realization (i.e. the portion of cells with morphological characteristics of apoptosis expressed in percentage terms, from the total amount of cells, expressing receptors of readiness to Fas-dependent apoptosis) was calculated. It has been found that the index of apoptosis realization was significantly decreased in alcoholic patients after the treatment in comparison with those before treatment (13.44% and 21.03% respectively).
Conclusion: Our data suggest that alcohol increases the readiness of lymphocytes to apoptosis and under the treatment the mechanisms controlling and limiting the processes of the programmed cell death are stimulated, hence the index of apoptosis realization significantly decreased.
Psychopharmacol. Biol. Narcol. 2005. Vol. 5, N 2. P. 910
Psyhopharmacology & biological narcology
ISSN 1606-8181
