Amitriptyline-induced loss of tight junction integrity in a human endothelial--smooth muscle cell bi-layer model.
Tricyclic antidepressants can, when taken in overdose, cause serious pulmonary failure such as the adult respiratory distress syndrome (ARDS). In this study we have examined the effects of some tricyclic antidepressants (amitriptyline, imipramine, nortriptyline and desipramine) on the viability and morphology of human endothelial and smooth muscle cells derived from umbilical cord. Effects of amitriptyline on endothelial cell fluidity, as well as permeability changes to an endothelial-smooth muscle cell bi-layer, were also studied. The tricyclic antidepressants induced acute, sub-lethal toxicity in both cell types above 100 microM as assessed by the MTT reduction assay. Morphological changes were also observed at these concentrations. Such changes were, however, absent at 33 microM and below. Amitriptyline did, however, cause a concentration-dependent fall in the electrical resistance of an endothelial-smooth muscle cell bi-layer, with significant effects already evident at 33 microM. All of these observed effects were fairly rapid and appeared within 5-15 min of exposure. The rapidity of these permeabilisation effects suggests potential membrane perturbations, since tricyclic antidepressants are lipophilic molecules with affinity for cell membranes. However, fluorescence anisotropy measurements showed no significant difference in membrane fluidity between amitriptyline-treated and control endothelial cells. Collectively, these data point to specific mechanisms of action of amitriptyline, and probably also the other tricyclic antidepressants studied, on endothelial permeability, which is a hallmark of ARDS. The data suggest that increased endothelial permeability could be due to impaired tight junction function.
Dahlin
K L
KL
Division of Inhalation Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Bohlin
K
K
Strindlund
J
J
Ryrfeldt
A
A
Cotgreave
I A
IA
D000639:Amitriptyline / Q000633:toxicity*; D000929:Antidepressive Agents, Tricyclic / Q000633:toxicity*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D003891:Desipramine / Q000633:toxicity; D017097:Electric Impedance; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000473:pathology; D019084:Fluorescent Antibody Technique, Indirect; D005562:Formazans / Q000378:metabolism; D006801:Humans; D007099:Imipramine / Q000633:toxicity; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000473:pathology; D009661:Nortriptyline / Q000633:toxicity; D013050:Spectrometry, Fluorescence; D013778:Tetrazolium Salts / Q000378:metabolism; D019108:Tight Junctions / Q000187:drug effects* / Q000502:physiology; D014470:Umbilical Cord / Q000166:cytology
D000929:Antidepressive Agents, Tricyclic; D005562:Formazans; D013778:Tetrazolium Salts; D000639:Amitriptyline; C079782:MTT formazan; D009661:Nortriptyline; D007099:Imipramine; D003891:Desipramine