title: Chlorambucil (nitrogen mustard) induced impairment of early vascular endothelial cell migration - effects of α-linolenic acid and N-acetylcysteine.
abstract: Alkylating agents (e.g. sulfur and nitrogen mustards) cause a variety of cell and tissue damage including wound healing disorder. Migration of endothelial cells is of utmost importance for effective wound healing. In this study we investigated the effects of chlorambucil (a nitrogen mustard) on early endothelial cells (EEC) with special focus on cell migration. Chlorambucil significantly inhibited migration of EEC in Boyden chamber and wound healing experiments. Cell migration is linked to cytoskeletal organization. We therefore investigated the distribution pattern of the Golgi apparatus as a marker of cell polarity. Cells are polarized under control conditions, whereas chlorambucil caused an encircling perinuclear position of the Golgi apparatus, indicating non-polarized cells. ROS are discussed to be involved in the pathophysiology of alkylating substances and are linked to cell migration and cell polarity. Therefore we investigated the influence of ROS-scavengers (α-linolenic acid (ALA) and N-acetylcysteine (NAC)) on the impaired EEC migration. Both substances, in particular ALA, improved EEC migration. Notably ALA restored cell polarity. Remarkably, investigations of ROS and RNS biomarkers (8-isoprostane and nitrotyrosine) did not reveal a significant increase after chlorambucil exposure when assessed 24h post exposure. A distinct breakdown of mitochondrial membrane potential (measured by TMRM) that recovered under ALA treatment was observed. In conclusion our results provide compelling evidence that the alkylating agent chlorambucil dramatically impairs directed cellular migration, which is accompanied by perturbations of cell polarity and mitochondrial membrane potential. ALA treatment was able to reconstitute cell polarity and to stabilize mitochondrial potential resulting in improved cell migration.
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany; Walther Straub Institute of Pharmacology and Toxicology, University of Munich, Goethestraße 33, 80336 Munich, Germany. Electronic address: dirk.steinritz@lrz.uni-muenchen.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Steinritz
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany; Department of Molecular and Cellular Sports Medicine, German Sports University, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany. Electronic address: annette2schmidt@bundeswehr.org.
authors.forename: Annette
authors.initials: A
authors.lastname: Schmidt
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany. Electronic address: thilo.simons@gmx.de.
authors.forename: Thilo
authors.initials: T
authors.lastname: Simons
authors.affiliation: Department of Molecular and Cellular Sports Medicine, German Sports University, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany. Electronic address: m.ibrahim@dshs-koeln.de.
authors.forename: Marwa
authors.initials: M
authors.lastname: Ibrahim
authors.affiliation: Department of Molecular and Cellular Sports Medicine, German Sports University, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany. Electronic address: morguet@mac.com.
authors.forename: Christian
authors.initials: C
authors.lastname: Morguet
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany. Electronic address: frankbalszuweit@bundeswehr.org.
authors.forename: Frank
authors.initials: F
authors.lastname: Balszuweit
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany. Electronic address: horstthiermann@bundeswehr.org.
authors.forename: Horst
authors.initials: H
authors.lastname: Thiermann
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstraße 11, 80937 Munich, Germany; Walther Straub Institute of Pharmacology and Toxicology, University of Munich, Goethestraße 33, 80336 Munich, Germany. Electronic address: kai.kehe@lrz.uni-muenchen.de.
authors.forename: Kai
authors.initials: K
authors.lastname: Kehe
authors.affiliation: Department of Molecular and Cellular Sports Medicine, German Sports University, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany. Electronic address: w.bloch@dshs-koeln.de.
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Bloch
authors.affiliation: Department of Molecular and Cellular Sports Medicine, German Sports University, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany. Electronic address: boelck@dshs-koeln.de.
authors.forename: Birgit
authors.initials: B
authors.lastname: Bölck
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000378:metabolism* / Q000633:toxicity; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology; D002699:Chlorambucil / Q000378:metabolism* / Q000633:toxicity; D042783:Endothelial Cells; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D007150:Immunohistochemistry; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism; D017962:alpha-Linolenic Acid / Q000494:pharmacology*
chemical_list: D018906:Antineoplastic Agents, Alkylating; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D017962:alpha-Linolenic Acid; D002699:Chlorambucil; D000111:Acetylcysteine
keywords: Chlorambucil; Endothelial cell; Migration; Mitochondrial membrane potential; Nitrogen mustard; ROS-scavenger