Login
Communauté Vinci
Extérieur
Si votre nom d'utilisateur ne se termine pas par @vinci.be ou @student.vinci.be, utilisez le formulaire ci-dessous pour accéder à votre compte de lecteur.
Titre : | The effect of Glyphosate and Atrazine on the Redox balance in the Beta pancreatic cells |
Auteurs : | Pierre-Loup Fretin, Auteur ; Leticia Prates Roma, Promoteur |
Type de document : | Travail de fin d'études |
Editeur : | Woluwe-Saint-Lambert : Haute École Léonard de Vinci, 2023 |
Langues: | Anglais |
Index. décimale : | TFE - Biologie médicale |
Mots-clés: | Atrazine ; Glyphosate ; cellule beta pancréatique ; stresse oxidatif |
Résumé : | In this study, our question is whether glyphosate and atrazine would induce oxidative stress in beta pancreatic cells. Glyphosate and atrazine are both pesticides that are known to affect human health and have been restricted or even banned in Europe but are still very much in use in other countries. Oxidative stress it's a state when the cells are in the imbalance of their redox situation. It can be detected by a high concentration of ROS, we will focus on one of the most common ROS detected in a state of oxidative stress. To be able to detect its presence, we will use a cell culture of rat beta-pancreatic(BRIN BD 11), which we will genetically manipulate to produce ROGFP, a fluorescent molecule known to be sensitive to oxidative molecules. We are going to add the ORP molecule, which will make the molecule more sensitive and specific to the presence of H2O2. We will then use two assays, one to detect this change in fluorescence when the cells are in contact with the pesticides and another one that will allow us to calculate the cytotoxicity of these pesticides when they are introduced into the cell environment. In the first, fluorescence assay, we have the objective of measuring the intensity in the two wavelengths of the spectrum of fluorescence of the ROGFP. And then we will calculate the ratio of fluorescence and the evolution of this ratio will allow us to know if the molecule is oxidised. If we see that there is an oxidation of this molecule, it will be an indication that there is a production of H2O2 in the cells when they are put in contact with the pesticides. In the results, we see no direct evidence that the pesticides are creating oxidative stress. The evolution of the ratio follows the example of the CTR medium, which is a stable situation, by slowly decreasing. Secondly, the MTT assay is a colorimetric assay, which will allow us to know if the cytotoxicity status/ cell proliferation is altered when pesticides are in contact with the cells in 24 hours. We will add a reactive, then if the cells are healthy they will be able to react with it and produce a colored product. We will read the absorbance in the wavelengths of the product color and then we will make a comparison between the absorbance and the number of cells and the cytotoxicity of the pesticide. For the atrazine, we see that in concentrations from 1 to 100 ppm, the cells are responding to the presence of the pesticide in a prolific way. When the concentration is at 500 ppm a drastic decrease in cell population and a high level of cytotoxicity. As for the glyphosate, we have a tendency in the cellular population to slowly decrease with the concentration. The cytotoxicity looks to be in negative values, which means that the cells are responding in a positive way, they fight back. Except at 500 ppm where the pesticides start to be toxic and kill the cells In the end, we cannot conclude that the pesticides cause a state of oxidative stress, but we can observe that at certain concentrations they appear to be toxic to beta pancreatic cells. |
Accès : | Identifiez-vous avant d'accéder au document électronique |
Disponible en ligne : | Oui |
Lieu du stage : | Allemagne, Hombourg CIPMM |
Département : | Biologie médicale |
Documents numériques (1)
Ce document n'est visible qu'après identification
TFE - Biologie médicale Adobe Acrobat PDF |