Effets des vitamines sur la fonction des lymphocytes.

Abstract

Résumé Afin de tester les effets des vitamines C et E in vitro sur des cellules en culture, nous avons travaillé sur un modèle de cellules humaines d’origine lymphocytaire. Les lymphocytes T sont isolées à partir d’un prélèvement sanguin et séparés par une centrifugation selon un gradient de densité de l’histopaque, puis incubés dans un milieu RPMI en présence d’un agent mitogène (la concanavaline A), des vitamines C et E à des concentrations différentes de 25 et 50 μM pendant 48 heures à 37°C et à 5% de CO2. Ces cellules sont par la suite utilisées pour analyser la prolifération cellulaire et le statut redox intracellulaire (vitamine C intra lymphocytaire, catalase, glutathion réduit, malondialdéhyde, protéines carbonylées). Nos résultats montrent que la concanavaline A stimule la prolifération des lymphocytes. Les vitamines C et E potentialisent cet effet d’une façon dose dépendante. En plus, la concanavaline A stimule la production du malondialdéhyde et des protéines carbonylées par les lymphocytes T en culture, avec une légère augmentation de la catalase. Les vitamines C et E régulent le stress oxydatif intracellulaire en diminuant les taux lymphocytaires en protéines carbonylées et en malondialdéhyde intracellulaires et en augmentant les défenses intracellulaires (vitamine C, GSH et catalase). De plus, l’effet protecteur de la vitamine C et E est plus important à la concentration de 50 μM. En conclusion, les vitamines C et E jouent un rôle immunostimulant et antioxydant ; la supplémentation en ces vitamines est donc à recommander. MotsMotsMotsMots-clés : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re: Vitamine C, vitamine E, cellule en culture, concanavaline A, statut re : Vitamine C, vitamine E, cellule en culture, concanavaline A, statut redox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, dox intracellulaire, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, lymphocyte T, catalase, catalase, malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéinemalondialdéhyde, glutathion réduit, protéine malondialdéhyde, glutathion réduit, protéines carbonylée carbonyléecarbonyléecarbonyléecarbonyléecarbonyléecarbonylée s. 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Abstract In order to test the in vitro effect of vitamins C and E on cells in culture, we have worked on a model of human cells of lymphocyte origin. T lymphocytes are isolated from blood sample and separated by centrifugation at a histopaque density gradient and then incubated in RPMI medium in the presence of a mitogen (concanavalin A), vitamins C And E at different concentrations of 25 and 50 μM for 48 hours at 37 °C and 5% CO2. These cells are subsequently used to analyze cell proliferation and intracellular redox status (vitamin C intra lymphocyte, catalase, reduced glutathione, malondialdehyde, carbonyl proteins). Our results show that concanavalin A stimulated lymphocyte proliferation. Vitamins C and E boosted this effect in a dose dependent manner. In addition, concanavalin A stimulated the production of malondialdehyde and carbonyl proteins by T lymphocytes in culture with a small increase in catalase activity. Vitamins C and E regulated intracellular oxidative stress by reducing lymphocyte levels of carbonyl proteins and malondialdehyde and by increasing intracellular antioxidant defense (vitamin C levels, GSH, catalase). In addition, the protective effect of vitamins was more pronounced at 50 μM. In conclusion, vitamins C and E displayed immune-stimulant and antioxidant effect; supplementation with these vitamins is then recommending. Keywords: vitamin C, vitamin E, culture cell, concanavalin A, intracellular redox status, T lymphocyte, catalase, malondialdehyde, reduced glutathione, carbonyl proteins. 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ملخص لدراسة تأثير الفيتامينات C و E في المخبر على زراعة الخلايا عملنا , على نموذج من أصل الخلية اللمفاوية البشرية. يتم عزل الخلية اللمفاوية التائية من عينة دم و فصل عن طريق الطرد المركزي و بالتدرج في كثافة Histopaque Histopaque HistopaqueHistopaque HistopaqueHistopaque و حضنت في وسط RPMI بوجود وكيل الفتيلي )كونكانافالين A ( و الفيتامينات C و E بتركيزات مختلفة ) 25 و 50 ( ميكرومول لمدة 48 ساعة عند 37 درجة مئوية و 5% CO2CO2 . ثم يتم استخدام هذه الخلايا لتحليل تكاثر الخلايا و حالة الأكسدة داخل الخلايا )فيتامين C داخل الخلية اللمفاوية , كاتالاز , ,glutathion,glutathion,glutathion,glutathion,glutathion ,glutathion ,glutathion réduitréduit réduitréduit Malondialdehyde Malondialdehyde MalondialdehydeMalondialdehydeMalondialdehyde Malondialdehyde MalondialdehydeMalondialdehydeMalondialdehyde و البروتين الكربونيل (. نتائجنا تظهر أن كونكانافالين A يحفزتكاثر الخلايا. الفيتامينات C و E تحفز بطريقة تعتمد على الجرعة و بالاضافة إلى ذلك , الكونكانافالين A تحفز انتاج ADM و البروتين الكربونيل من الخلايا اللمفاوية التائية المزرعة مع زيادة طفيفة من الكاتالاز. الفيتامينات C و E تنظم الأكسدة داخل الخلايا عن طريق خفض مستويات الخلايا اللمفاوية في البروتين الكربونيل داخل الخلايا و كذلك DAM و زيادة دفاع الخلايا )فيتامين C , GSH, والكاتالاز ( و بالاضافة الى ذلك, فان تأثير الوقائي للفيتامين C و E يكون أكبر بتركيز 50 ميكرومول. وفي الختام, فان الفيتامينات C و E تلعب دور في تحفيز المناعة و كذلك مضاد للأكسدة و يوصى بها كمكمل غذائي. الكلمات المفتاحية : فيتامين C , فيتامين E , زراعة الخلايا, كونكانافالين A , حالة الاكسدة داخل الخلايا, اللمفاوية التائية, كاتالاز, MDA MDA , GSHGSH , البروتين الكربونيل