Volatile Anesthetics Induce Caspase-dependent, Mitochondria-mediated Apoptosis in Human T Lymphocytes In Vitro |
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Authors: | Loop, Torsten M.D. Dovi-Akue, David Cand. Med. Frick, Michael Cand. Med. Roesslein, Martin M.D. Egger, Lotti Ph.D.
Humar, Matjaz Ph.D.
Hoetzel, Alexander M.D. Schmidt, Rene M.D. Borner, Christoph Ph.D. Pahl, Heike L. Ph.D.# Geiger, Klaus K. M.D. Pannen, Benedikt H. J. M.D. |
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Affiliation: | Loop, Torsten M.D.*; Dovi-Akue, David Cand. Med.†; Frick, Michael Cand. Med.†; Roesslein, Martin M.D.‡; Egger, Lotti Ph.D.§; Humar, Matjaz Ph.D.§; Hoetzel, Alexander M.D.‡; Schmidt, Rene M.D.‡; Borner, Christoph Ph.D.∥; Pahl, Heike L. Ph.D.#; Geiger, Klaus K. M.D.**; Pannen, Benedikt H. J. M.D.†† |
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Abstract: | Background: Volatile anesthetics modulate lymphocyte function during surgery, and this compromises postoperative immune competence. The current work was undertaken to examine whether volatile anesthetics induce apoptosis in human T lymphocytes and what apoptotic signaling pathway might be used. Methods: Effects of sevoflurane, isoflurane, and desflurane were studied in primary human CD3+ T lymphocytes and Jurkat T cells in vitro. Apoptosis and mitochondrial membrane potential were assessed using flow cytometry after green fluorescent protein-annexin V and DiOC6-fluorochrome staining. Activity and proteolytic processing of caspase 3 was measured by cleaving of the fluorogenic effector caspase substrate Ac-DEVD-AMC and by anti-caspase-3 Western blotting. Release of mitochondrial cytochrome c was studied after cell fractionation using anti-cytochrome c Western blotting and enzyme-linked immunosorbent assays. Results: Sevoflurane and isoflurane induced apoptosis in human T lymphocytes in a dose-dependent manner. By contrast, desflurane did not exert any proapoptotic effects. The apoptotic signaling pathway used by sevoflurane involved disruption of the mitochondrial membrane potential and release of cytochrome c from mitochondria to the cytosol. In addition, the authors observed a proteolytic cleavage of the inactive p32 procaspase 3 to the active p17 fragment, increased caspase-3-like activity, and cleavage of the caspase-3 substrate poly-ADP-ribose-polymerase. Sevoflurane-induced apoptosis was blocked by the general caspase inhibitor Z-VAD.fmk. Death signaling was not mediated via the Fas/CD95 receptor pathway because neither anti-Fas/CD95 receptor antagonism nor FADD deficiency or caspase-8 deficiency were able to attenuate sevoflurane-mediated apoptosis. |
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