Agomelatine’s effect on human genetic material: in vitro study

Εισαγωγή: Η αγομελατίνη αποτελεί ένα εγκεκριμένο φάρμακο για τη θεραπεία της μείζονος καταθλιπτικής διαταραχής. Παρουσιάζει δράση μελατονινεργικού αγωνιστή και ανταγωνιστή των 5-HT2C υποδοχέων της σεροτονίνης. Σκοπός της παρούσας έρευνας είναι η μελέτη της επίδρασης της αγομελατίνης στο ανθρώπινο DNA in vitro με τον υπολογισμό ευαίσθητων κυτταρογενετικών δεικτών.Υλικό και Μέθοδος: Oι χρωματιδιακές ανταλλαγές (Sister Chromatid Exchanges, SCEs) θεωρούνται ένας από τους πιο ευαίσθητους δείκτες γονοτοξικότητας. O δείκτης ρυθμού πολλαπλασιασμού (Proliferation Rate Index, PRI) είναι ένας από τους πιο αξιόπιστους δείκτες κυτταροστατικότητας, ενώ ο μιτωτικός δείκτης (Mitotic Index, MI) δείχνει με ακρίβεια την ικανότητα του κυττάρου για πολλαπλασιασμό. Αρχικά παρασκευάστηκαν διαλύματα αγομελατίνης πέντε διαφορετικών συγκεντρώσεων (A=2.5μg/ml, B=5μg/ml, C=10μg/ml, D=15μg/ml και E=20μg/ml). Oι συγκεντρώσεις B και C είναι οι πιο συχνά χρησιμοποιούμενες στην κλινική πράξη. Τα διαλύματα προστέθηκαν σε καλλιέργειες λεμφοκυττάρων από περιφερικό αίμα τεσσάρων νεαρών υγειών αιμοδοτών. Μετά από 72 ώρες επώασης, με την κατάλληλη τεχνική τα καλλιεργημένα λεμφοκύτταρα επιστρώθηκαν σε αντικειμενοφόρους πλάκες, χρωματίστηκαν με την μέθοδο Fluorescence plus Giemsa και οι προαναφερθέντες δείκτες υπολογίστηκαν με οπτικό μικροσκόπιο.Αποτελέσματα: Η ανάλυση των αποτελεσμάτων αποκάλυψε στατιστικά σημαντική αύξηση των χρωματιδιακών ανταλλαγών και μείωση τόσο του δείκτη ρυθμού πολλαπλασιασμού όσο και του μιτωτικού δείκτη . Επιπρόσθετα, προέκυψε συσχέτιση μεταξύ α) της αύξησης των SCEs και των μεταβολών του ΜΙ και του PRI και β) των μεταβολών ΜΙ-PRI.Συμπεράσματα: Η αγομελατίνη σε θεραπευτικές δόσεις προκάλεσε δοσοεξαρτώμενες μεταβολές στους υπό μελέτη κυτταρογενετικούς δείκτες. Το παραπάνω ενδέχεται να καταδεικνύει στοιχεία για το μηχανισμό δράσης του φαρμάκου. Λαμβάνοντας υπόψη την αυξανόμενη χρήση της αγομελατίνης επιβάλλεται περεταίρω μελέτη της κυτταρογενετικής της δράσης σε περισσότερους αιμοδότες καθώς και σε άλλες κυτταρικές σειρές.Introduction: Agomelatine is a prescription drug approved for the treatment of major depressive disorder. It is a melatonergic agonist and a 5-HT2C antagonist. The cytogenetic behavior of agomelatine has not been studied. The aim of the present study is the investigation of the in vitro effect of agomelatine on human DNA, by estimating sensitive cytogenetic indices. Methods: SCEs (Sister Chromatid Exchanges) are considered as one of the most sensitive markers of genotoxicity, PRI (Proliferation Rate Index) is one of the most reliable markers of cytostatic activity, whereas MI (Mitotic Index) shows precisely the ability of the cell to proliferate. We have investigated the effect of five agomelatine solutions on SCEs, PRI and MI of human cultured lymphocytes stained with the Fluorescence plus Giemsa method and estimated with the optical microscope. Results: Analysis of the results has revealed statistically significant (p<0.001) dose-dependent increase of SCE frequencies and significant reduction of PRI and MI values on lymphocyte cultures treated with agomelatine. Furthermore, a correlation was observed between a) the magnitude of the SCE induction and the PRI alterations, b) the magnitude of the MI alterations and the SCE induction and c) the magnitude of PRI alterations and MI alterations. Conclusions:Agomelatine at therapeutic doses exhibited dose-dependent cytogenetic activity in vitro. This may provide additional information about the mechanism of action of the drug. Considering that the use of agomelatine has rapidly increased, further studies in other cell lines and in vivo experimental settings are needed in order to evaluate its effect on human genetic material

Venlafaxine’s effect on human genetic material: in vitro study

Introduction: Venlafaxine is a prescription drug approved for the treatment of major depressive disorder, generalized anxiety disorder, panic disorder and social anxiety disorder.  It is a serotonin and norepinephrine reuptake inhibitor and a weak dopamine reuptake inhibitor. The aim of the present study is to investigate the in vitro effect of venlafaxine on human genetic material, by estimating sensitive cytogenetic indices.Methods: Five venlafaxine solutions (A=15μg/ml, B=30μg/ml, C=45μg/ml, D=60μg/ml, E=75μg/ml) were added to cultures of peripheral blood lymphocytes of six healthy donors. After 72 hours of incubation, the cultured lymphocytes were plated on glass slides, stained with the Fluorescence plus Giemsa method and Sister Chromatid Exchanges (SCEs), a sensitive marker of genotoxicity, Proliferation Rate Index (PRI), a reliable marker of cytostatic activity and Mitotic Index (MI), a marker which shows precisely the ability of a cell to proliferate were measured with the optical microscope.Results: Result analysis revealed t: a) a statistically significant (p=0.001) dose-dependent increase of SCEs and b) a statistically significant (p=0.001) reduction of PRI and MI in all concentrations. Furthermore, a correlation was observed between a) SCE and PRI index variations, b) MI and SCE index variations and c) PRI and MI index variations. Conclusions: Venlafaxine exhibited dose-dependent cytogenetic activity in vitro, increasing SCE frequencies and diminishing PRI and MI levels in healthy human cultured lymphocytes. Venlafaxine as other antidepressants seems to affect human T lymphocytes by modifying epigenetic and DNA replication procedures. This may provide additional information about the mechanism of action of the drug. Considering that the use of venlafaxine has rapidly increased with many off label indications, further studies in other cell lines and in vivo experimental settings are needed in order to evaluate its potential effects on human genetic material

The effect of 3,4-methylenedioxymethamphetamine (MDMA) on human genetic material: an in vitro study

Background: 3,4-methylenedioxymethamphetamine (MDMA), is a synthetic illicit psychostimulant drug  that affects  mood and social interactions. The aim of the present study is to investigate the in vitro effect of MDMA on human genetic material, by estimating sensitive cytogenetic indices.Methods: MDMA solutions (A=15μg/ml, B=30μg/ml, C=45μg/ml, D=60μg/ml, E=75μg/ml) were added in cultures of peripheral blood lymphocytes of four healthy donors. After 72 hours of incubation, the cultured lymphocytes were collected, plated on glass slides, stained with the Fluorescence plus Giemsa method and SCEs, PRI and MI were measured with the optical microscope. Sister Chromatid Exchanges (SCEs) is a sensitive marker of genotoxicity, Proliferation Rate Index (PRI) is a reliable marker of cytostatic activity and Mitotic Index (MI) is a reliable indicator of cell ability to proliferate.Results: Result analysis revealed: a) a statistically significant (p=0.001) reduction of SCEs on lower MDMA concentration and a significant induction (p=0.001) of SCEs after the effect of higher MDMA concentrations, b) PRI and MI reduction (p=0.001) after the effect of MDMA concentrations 45, 60 and 75μg/ml. Correlation was observed between a) SCE and PRI index variations, b) MI and SCE index variations and c) PRI and MI index variations. Conclusions: MDMA exhibited an interesting cytogenetic activity in vitro. It seems to affect human T lymphocytes by epigenetic and DNA replication modifications. This may provide additional information about the mechanism of action of the drug. Further studies in other cell lines and in vivo experimental settings are needed in order to evaluate its potential effects on human genetic material