Effects of nitric oxide on proliferation and differentiation of rat brown adipocytes in primary cultures
Effects of nitric oxide on proliferation and differentiation of rat brown adipocytes in primary cultures
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DOI：
10.1038/sj.bjp.0702131
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发表时间：
1998-10-01
影响因子：
7.3
通讯作者：
Carruba, MO
中科院分区：
文献类型：
--
作者：
Nisoli, E;Clementi, E;Carruba, MO
1 在本工作中，我们研究了 NO 对原代培养物中棕色脂肪细胞增殖和分化的影响。2 从大鼠棕色脂肪组织中分离出的棕色脂肪前体细胞培养 8 天直至汇合，并每天用 NO 供体试剂 S-亚硝基-乙酰青霉胺 (SNAP) 或 S-亚硝基-L-谷胱甘肽 (GSNO) 进行处理。两种药物 (300 μM) 在第 8 天均使细胞增殖减少约 8 倍。NO 的抑制作用不太可能是由于细胞毒性，因为 (i) 即使在重复添加 SNAP 或 GSNO 8 天后，细胞也从未完全失去其增殖能力，并且 (ii) 除去含有 NO 供体的培养基后，抑制作用是可逆的。3 每日用一氧化氮合酶抑制剂（例如 NG-硝基-L-精氨酸甲酯）进行治疗（L-NAME，300 mu M），导致细胞增殖刺激 44 +/- 5%，n = 3，表明棕色脂肪细胞内源性产生的 NO 可能参与调节细胞生长。4 每日用 SNAP 或 GSNO 治疗可诱导显着的线粒体生成，测量为 3-[4,5-二甲基噻唑-2-基-]-2,5-二苯基四唑的线粒体转化溴化物 (MTT) 转化为甲臜，而每天用 L-NAME 治疗则没有效果。5 NO 供体对细胞增殖的抑制伴随着编码过氧化物酶体增殖物激活受体-γ 和解偶联蛋白-1 的两个基因的表达，这两个基因在分化过程中上调。6 通过 8-溴环 GMP (100-1000 mu M) 增加细胞中的环 GMP 并没有重现观察到的 NO 对细胞数量或基因表达的影响。另一方面，用 NO 刺激的鸟苷酸环化酶抑制剂 1H-[1,2,4]恶二唑[4,3-a]喹喔啉-1-酮 (ODQ) 进行长期治疗，可减少过氧化物酶体增殖物激活受体-γ 和解偶联蛋白-1 的表达。
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 mu M) 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 NG-nitro-L-arginine methyl ester (L-NAME, 300 mu M), 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-gamma and uncoupling protein-1, which are upregulated during differentiation.6 Increasing cyclic GMP in cells by 8-bromo-cyclic GMP (100-1000 mu M) 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 receptor-gamma and uncoupling protein-1.