OBJECTIVE: To elucidate the effects and molecular mechanism(s) by means of which noradrenaline (NA) protects against the tumor necrosis factor (TNF)-α-induced apoptosis of brown adipocytes. DESIGN: Brown fat precursor cells were isolated from young rats; 2.5 million cells were added to each 24-well culture plate and cultured in a defined culture medium. On day 8, the cultured cells were exposed to murine recombinant TNF-α and/or cycloheximide (CHX; 10 μg/ml) and/or NA and/or nitric oxide (NO) donors and/or the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and/or 10 μM heat shock protein 70 (HSP70) antisense or sense oligomers. MEASUREMENTS: Analysis of DNA fragmentation on agarose gel as a marker of apoptosis; reverse transcriptase-polymerase chain reaction analysis of mRNA levels; Western blotting analysis of protein levels. RESULTS: Pretreatment of primary cultures of rat brown fat cells with micromolar concentrations of NA or the NO-donor S-nitroso-N-acetylpenicillamine (SNAP) induced the expression of HSP70 mRNA and protein, which was associated with cytoprotection against TNF-α plus CHX-induced apoptosis. The L-NAME inhibitor of NO synthase activity inhibited both NA-stimulated HSP70 expression and cytoprotection. Furthermore, pretreatment of brown adipocytes with an antisense oligonucleotide to HSP70 antagonized both SNAP- and NA-induced cytoprotection. CONCLUSION: These findings demonstrate that the NO produced by NA stimulation can induce resistance to the TNF-α-induced apoptosis of brown adipocytes, possibly by means of the expression of HSP70.

Nisoli, E., Regianini, L., Bulbarelli, A., Briscini, L., Breacale, R., & Carruba, M. (2001). Protective effects of noradrenaline against tumor necrosis factor-α-induced apoptosis in cultured rat brown adipocytes: Role of nitric oxide-induced heat shock protein 70 expression. INTERNATIONAL JOURNAL OF OBESITY, 25(10), 1421-1430 [10.1038/sj.ijo.0801788].

Protective effects of noradrenaline against tumor necrosis factor-α-induced apoptosis in cultured rat brown adipocytes: Role of nitric oxide-induced heat shock protein 70 expression

Bulbarelli, A.;
2001

Abstract

OBJECTIVE: To elucidate the effects and molecular mechanism(s) by means of which noradrenaline (NA) protects against the tumor necrosis factor (TNF)-α-induced apoptosis of brown adipocytes. DESIGN: Brown fat precursor cells were isolated from young rats; 2.5 million cells were added to each 24-well culture plate and cultured in a defined culture medium. On day 8, the cultured cells were exposed to murine recombinant TNF-α and/or cycloheximide (CHX; 10 μg/ml) and/or NA and/or nitric oxide (NO) donors and/or the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and/or 10 μM heat shock protein 70 (HSP70) antisense or sense oligomers. MEASUREMENTS: Analysis of DNA fragmentation on agarose gel as a marker of apoptosis; reverse transcriptase-polymerase chain reaction analysis of mRNA levels; Western blotting analysis of protein levels. RESULTS: Pretreatment of primary cultures of rat brown fat cells with micromolar concentrations of NA or the NO-donor S-nitroso-N-acetylpenicillamine (SNAP) induced the expression of HSP70 mRNA and protein, which was associated with cytoprotection against TNF-α plus CHX-induced apoptosis. The L-NAME inhibitor of NO synthase activity inhibited both NA-stimulated HSP70 expression and cytoprotection. Furthermore, pretreatment of brown adipocytes with an antisense oligonucleotide to HSP70 antagonized both SNAP- and NA-induced cytoprotection. CONCLUSION: These findings demonstrate that the NO produced by NA stimulation can induce resistance to the TNF-α-induced apoptosis of brown adipocytes, possibly by means of the expression of HSP70.
Articolo in rivista - Articolo scientifico
Scientifica
Apoptosis; Brown adipocytes; Heat shock protein; Nitric oxide; Noradrenaline; Adipose Tissue, Brown; Animals; Apoptosis; Blotting, Western; Cells, Cultured; Cycloheximide; DNA Fragmentation; Electrophoresis, Agar Gel; HSP70 Heat-Shock Proteins; Male; Nitric Oxide; Nitroso Compounds; Norepinephrine; Oligonucleotides, Antisense; Protein Synthesis Inhibitors; RNA, Messenger; Rats; Rats, Zucker; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha; Medicine (miscellaneous); Public Health, Environmental and Occupational Health; Endocrinology; Food Science; Endocrinology, Diabetes and Metabolism
English
Nisoli, E., Regianini, L., Bulbarelli, A., Briscini, L., Breacale, R., & Carruba, M. (2001). Protective effects of noradrenaline against tumor necrosis factor-α-induced apoptosis in cultured rat brown adipocytes: Role of nitric oxide-induced heat shock protein 70 expression. INTERNATIONAL JOURNAL OF OBESITY, 25(10), 1421-1430 [10.1038/sj.ijo.0801788].
Nisoli, E; Regianini, L; Bulbarelli, A; Briscini, L; Breacale, R; Carruba, M
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/10281/187352
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