Preincubation with atrial natriuretic peptide protects NG108-15 cells against the toxic/proapoptotic effects of the nitric oxide donor S-nitroso-N-acetylpenicillamine
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AbstractThe TUNEL method is used to quantify the proapoptotic effects of an NO donor, S-nitroso-N-acetylpenicillamine (SNAP), in NG108-15 cells. Unlike sodium nitroprusside used in previous studies, SNAP does not release cyanide along with NO, thus NO toxicity was determined without concurrent cyanide toxicity. The present study also determined if pretreatment with ANP could protect against NO-induced apoptosis in NG108-15 cells. Cell death at 24 h following SNAP treatment was associated with apoptotic DNA fragmentation. SNAP at 0.5, 0.75, 1.0, and 2.0 mM caused significant (P<0.05) increases in the percentage of TUNEL-labeled cells from a control of 0.90% to 6.19%, 6.36%, 7.25%, and 15.1%, respectively. Thus, SNAP caused concentration-dependent induction of apoptosis in NG108-15 cells. SNAP-induced apoptosis was confirmed by morphological changes and increased levels of polynucleosome-sized fragments of DNA assessed by capillary electrophoresis. Preincubation for 24 h with ANP at 0.01, 0.1, and 1.0 muM, before the SNAP, significantly (P<0.05) decreased the percentage of labeled cells from 7.25% to 5.10%, 4.36%, and 3.24% in the presence of SNAP (1 mM) and from 15.1% to 7.91%, 6.64%, and 5.60% in the presence of SNAP (2 mM), respectively, representing protection of 24.0%, 34.0%, and 57.0% against SNAP (1 mM) and 26.0%, 37.0%, and 50.9% against SNAP (2 mM). Thus, prior activation of a cGMP-mediated neuroprotective mechanism induced by ANP appears to counterbalance, at least partially, the proapoptotic effects of excess NO. This neuroprotective mechanism involving cGMP may be especially important in protecting against the development of neurodegenerative diseases in which excess NO is thought to contribute to neuronal apoptosis.
All Author(s) ListChew SBC, Leung PY, Fiscus RR
Journal nameHistochemistry and Cell Biology
Volume Number120
Issue Number3
Pages163 - 171
LanguagesEnglish-United Kingdom
KeywordsANP; apoptosis; neural cells; SNAP; TUNEL
Web of Science Subject CategoriesCell Biology; CELL BIOLOGY; Microscopy; MICROSCOPY

Last updated on 2021-13-01 at 00:44