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2017 - Trifluoperazine-Induced Suicidal Erythrocyte Death and S-Nitrosylation Inhibition, Reversed by the Nitric Oxide Donor Sodium Nitroprusside

icon pl   Patchliner publication in Cellular Physiology and Biochemistry (2017)

Ghashghaeinia M., Wesseling M.C., Ramos E., Petkova-Kirova P., Waibel S., Lang E., Bissinger R., Alzoubi K., Edelmann B., Hosseinzadeh Z., Dreischer P., Shahvaroughi-Farahani A., Mrowietz U., Köberle M., Kaestner L., Bernhardt I., Martínez-Ruiz A., Wieder T., Lang F.


Cellular Physiology and Biochemistry (2017) 42:1985–1998


Background and Purpose:
The high potency antipsychotic drug trifluoperazine (10-[3-(4-methyl-1-piperazinyl)-propyl]-2-(trifluoromethyl)-(10)H-phenothiazine dihydrochloride; TFP) may either counteract or promote suicidal cell death or apoptosis. Similar to apoptosis, erythrocytes may enter eryptosis, characterized by phosphatidylserine exposure at the cell surface and cell shrinkage. Eryptosis can be stimulated by an increase in cytoplasmic Ca2+ concentration ([Ca2+]i) and inhibited by nitric oxide (NO). We explored whether TFP treatment of erythrocytes induces phosphatidylserine exposure, cell shrinkage, and calcium influx, whether it impairs S-nitrosylation and whether these effects are inhibited by NO.

Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, and protein nitrosylation from fluorescence switch of the Bodipy-TMR/Sypro Ruby signal.

Exposure of human erythrocytes to TFP significantly enhanced the percentage of annexin-V-binding cells, raised [Ca2+]i, and decreased S-nitrosylation. The effect of TFP on annexin-V-binding was not affected by removal of extracellular Ca2+ alone, but was significantly inhibited by pre-treatment with sodium nitroprusside (SNP), an effect significantly augmented by additional removal of extracellular Ca2+. A 3 hours treatment with 0.1 µM Ca2+ ionophore ionomycin triggered annexin-V-binding and cell shrinkage, effects fully reversed by removal of extracellular Ca2+. Conclusions: TFP induces eryptosis and decreases protein S-nitrosylation, effects blunted by nitroprusside. The effect of nitroprusside is attenuated in the presence of extracellular Ca2+.

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Ghashghaeinia M.a,b· Wesseling M.C.c· Ramos E.d,e· Petkova-Kirova P.f· Waibel S.b· Lang E.b·Bissinger R.b· Alzoubi K.b· Edelmann B.g· Hosseinzadeh Z.h· Dreischer P.i· Shahvaroughi-Farahani A.j· Mrowietz U.a· Köberle M.k· Kaestner L.f,l· Bernhardt I.c· Martínez-Ruiz A.d,e·Wieder T.m· Lang F.n

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