TY - JOUR
T1 - Effects of nitric oxide on proliferation and differentiation of rat brown adipocytes in primary cultures
AU - Nisoli, Enzo
AU - Clementi, Emilio
AU - Tonello, Cristina
AU - Sciorati, Clara
AU - Briscini, Luca
AU - Carruba, Michele O.
PY - 1998
Y1 - 1998
N2 - 1. In the present work, we study the effect of NO on the proliferation and differentiation of brown fat cells in primary cultures. 2. Brown fat precursor cells isolated from rat brown adipose tissue were cultured for 8 days until confluence and treated daily with the NO donating agents, S-nitroso-acetyl penicillamine (SNAP or S-nitroso-L-glutathione (GSNO). Both agents (300 μM) decreased cell proliferation approximately 8 fold on day 8. The inhibitory effect of NO was unlikely to be due to cytotoxicity since (i) cells never completely lost their proliferation capacity even after 8 days of exposure to repeated additions of SNAP or GSNO, and (ii) the inhibitory effect was reversible after removal of the media containing NO donors. 3. Daily treatment with nitric oxide synthase inhibitors, such as N(G)-nitro-L-arginine methyl ester (L-NAME, 300 μM), led to the stimulation of cell proliferation by 44 ± 5%, n = 3, suggesting that NO, endogenously produced in brown adipocytes, may be involved in modulating cell growth. 4. Daily treatment with both SNAP or GSNO induced significant mitochondriogenesis, measured as the mitochondrial conversion of 3-[4,5-dimethylthiazol-2-yl-]-2,5-diphenyl tetrazolium bromide (MTT) to formazan, whilst daily treatment with L-NAME was without effect. 5. The inhibition of cell proliferation by NO donors was accompanied by the expression of two genes coding for peroxisome proliferator activated receptor-γ and uncoupling protein-1, which are upregulated during differentiation. 6. increasing cyclic GMP in cells by 8-bromo-cyclic GMP (100-1000 μM) did not reproduce the observed NO effects on either cell number or gene expression. On the other hand, chronic treatment with the inhibitor of the NO-stimulated guanylyl cyclase, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ), reduced the expression of peroxisome proliferator activated receplor-γ and uncoupling protein-1.
AB - 1. In the present work, we study the effect of NO on the proliferation and differentiation of brown fat cells in primary cultures. 2. Brown fat precursor cells isolated from rat brown adipose tissue were cultured for 8 days until confluence and treated daily with the NO donating agents, S-nitroso-acetyl penicillamine (SNAP or S-nitroso-L-glutathione (GSNO). Both agents (300 μM) decreased cell proliferation approximately 8 fold on day 8. The inhibitory effect of NO was unlikely to be due to cytotoxicity since (i) cells never completely lost their proliferation capacity even after 8 days of exposure to repeated additions of SNAP or GSNO, and (ii) the inhibitory effect was reversible after removal of the media containing NO donors. 3. Daily treatment with nitric oxide synthase inhibitors, such as N(G)-nitro-L-arginine methyl ester (L-NAME, 300 μM), led to the stimulation of cell proliferation by 44 ± 5%, n = 3, suggesting that NO, endogenously produced in brown adipocytes, may be involved in modulating cell growth. 4. Daily treatment with both SNAP or GSNO induced significant mitochondriogenesis, measured as the mitochondrial conversion of 3-[4,5-dimethylthiazol-2-yl-]-2,5-diphenyl tetrazolium bromide (MTT) to formazan, whilst daily treatment with L-NAME was without effect. 5. The inhibition of cell proliferation by NO donors was accompanied by the expression of two genes coding for peroxisome proliferator activated receptor-γ and uncoupling protein-1, which are upregulated during differentiation. 6. increasing cyclic GMP in cells by 8-bromo-cyclic GMP (100-1000 μM) did not reproduce the observed NO effects on either cell number or gene expression. On the other hand, chronic treatment with the inhibitor of the NO-stimulated guanylyl cyclase, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ), reduced the expression of peroxisome proliferator activated receplor-γ and uncoupling protein-1.
KW - Brown adipocytes
KW - Cell differentiation
KW - Cell proliferation
KW - Energy expenditure
KW - Nitric oxide
KW - Obesity
KW - Peroxisome proliferator-activated receptor-γ
KW - Uncoupling protein-1
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U2 - 10.1038/sj.bjp.0702131
DO - 10.1038/sj.bjp.0702131
M3 - Article
C2 - 9831929
AN - SCOPUS:0031727870
VL - 125
SP - 888
EP - 894
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
SN - 0007-1188
IS - 4
ER -