Tricyclic antidepressant amitriptyline attenuates Ca2+ responses in rat peritoneal macrophages

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Рұқсат жабық Тек жазылушылар үшін

Аннотация

Amitriptyline is a tricyclic antidepressant widely used in clinical practice for the treatment of anxiety, depression and chronic pain. These drugs have a multifaceted effect on cellular processes. One of their targets is sigma-1 receptors. Sigma-1 receptors are molecular chaperones located in endoplasmic reticulum membrane; they are characterized by a unique structure and pharmacological profile. Sigma-1 receptors regulate many cellular processes in health and disease, including Ca2+ signaling. Using Fura-2AM microfluorimetry, it was shown for the first time that sigma-1 receptor agonist, antidepressant amitriptyline, significantly suppresses both Ca2+ mobilization from intracellular Ca2+-stores and subsequent store-dependent Ca2+ entry into cells, induced by endoplasmic Ca2+-ATPase inhibitors thapsigargin and cyclopiazonic acid, as well as disulfide-containing immunomodulators glutoxim and molixan, in rat peritoneal macrophages. The results suggest the involvement of sigma-1 receptors in a complex signaling cascade induced by glutoxim or molixan, leading to an increase of intracellular Ca2+ concentration in macrophages. The results also indicate the participation of sigma-1 receptors in the regulation of store-dependent Ca2+ entry in macrophages.

Толық мәтін

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Авторлар туралы

L. Milenina

St. Petersburg State University

Хат алмасуға жауапты Автор.
Email: l.milenina@spbu.ru

Chair of Biophysics

Ресей, St. Petersburg, 199034

Z. Krutetskaya

St. Petersburg State University

Email: z.krutetskaya@spbu.ru

Chair of Biophysics

Ресей, St. Petersburg, 199034

V. Antonov

St. Petersburg State Pediatric Medical University

Email: l.milenina@spbu.ru

Chair of Biochemistry

Ресей, St. Petersburg, 194100

N. Krutetskaya

St. Petersburg State University

Email: l.milenina@spbu.ru

Chair of Biophysics

Ресей, St. Petersburg, 199034

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2. Fig. 1. Effect of amitriptyline on the increase in [Ca2+]i caused by glutoxim in rat peritoneal macrophages. Here and in Fig. 2–4 on the ordinate - the ratio of fluorescence intensities of Fura-2AM at exciting wavelengths of 340 and 380 nm (F340/F380); a – macrophages were incubated for 20 min in the presence of 100 μg/ml glutoxim in a nominally calcium-free medium, then Ca2+ was initiated by introducing 2 mM Ca2+ into the external environment; against the background of the developed Ca2+ influx, 40 μg/ml amitriptyline was added; b – macrophages located in a calcium-free medium were incubated for 20 min with 20 μg/ml amitriptyline, then 100 μg/ml glutoxim was added, after 20 min, Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium. Here and in Fig. 2–4 each recording was obtained for a group of 40–50 cells and represents a typical version of 6–8 independent experiments.

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3. Fig. 2. Effect of amitriptyline on the increase in [Ca2+]i caused by molixan in rat peritoneal macrophages: a – macrophages were incubated for 20 min in the presence of 100 μg/ml molixan in a nominally calcium-free medium, then Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium; against the background of the developed Ca2+ influx, 40 μg/ml amitriptyline was added; b – macrophages located in a calcium-free medium were incubated for 20 min with 20 μg/ml amitriptyline, then 100 μg/ml molixan was added, after 20 min, Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium.

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4. Fig. 3. Effect of amitriptyline on Ca2+ responses induced by thapsigargin in rat peritoneal macrophages: a – macrophages were stimulated with 0.5 μM thapsigargin in a nominally calcium-free medium, then Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external environment; against the background of the developed Ca2+ influx, 40 μg/ml amitriptyline was added; b – macrophages were pre-incubated for 20 min with 20 µg/ml amitriptyline in a calcium-free medium, then 0.5 µM thapsigargin was added, after which Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium.

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5. Fig. 4. Effect of amitriptyline on Ca2+ responses induced by cyclopiazonic acid (CPA) in rat peritoneal macrophages: a – macrophages were stimulated with 10 μM CPA in a nominally calcium-free medium, then Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium; against the background of the developed Ca2+ influx, 40 μg/ml amitriptyline was added; b – macrophages were pre-incubated for 20 min with 20 µg/ml amitriptyline in a calcium-free medium, then 10 µM CPA was added, after which Ca2+ entry was initiated by introducing 2 mM Ca2+ into the external medium.

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