Nitric oxide production by the vacuolar-type (H+)-ATPase inhibitors bafilomycin A1 and concanamycin a and its possible role in apoptosis in RAW 264.7 cells
Nitric oxide production by the vacuolar-type (H+)-ATPase inhibitors bafilomycin A1 and concanamycin a and its possible role in apoptosis in RAW 264.7 cells
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DOI：
10.1124/jpet.106.109280
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发表时间：
2006-11-01
影响因子：
3.5
通讯作者：
Ohuchi, Kazuo
中科院分区：
文献类型：
--
作者：
Hong, JangJa;Nakano, Yasuhiro;Ohuchi, Kazuo
在小鼠白血病单核细胞RAW 264.7中，液泡型(H+)- atp酶（v - atp酶）抑制剂巴filomycin A1和concanamycin A通过表达诱导型一氧化氮合成酶mRNA及其蛋白诱导一氧化氮（NO）的产生，并通过3-(4,5-二甲基（噻唑-2-基）-2,5-二苯基溴化四唑（MTT）测定降低细胞生长和存活。巴霉素A1和康纳霉素A激活核因子(NF)- κ B和激活蛋白-1，降低κ B- α水平，升高磷酸化的c-Jun n-末端激酶（JNK）水平。这些V-ATPase抑制剂诱导的NO生成被NF-kappa B抑制剂Bay 11-7082 [(E)3-[（4-甲基苯基）磺酰基])2-丙烯腈]和JNK抑制剂SP600125[炭黑[1,9cd]吡唑-6(2H)- 1]抑制，同时部分减轻V-ATPase抑制剂诱导的MTT反应下降。Na+， K+- atp酶抑制剂二布卡因和f - atp酶抑制剂寡霉素不诱导NO的产生，其浓度降低了MTT反应。NO供体S-nitrosoN- acetyl-DL-penicillamine进一步降低了V-ATPase抑制剂诱导的MTT反应的降低，NO清除剂2-（4-羧基苯基）-4,4,5,5-四甲基咪唑啉- 1-氧-3-氧化物钠盐（羧基- ptio）部分缓解了这一现象。用一氧化氮合酶抑制剂n - g - monom甲基- l-精氨酸醋酸酯处理后，巴霉素A1和康那霉素A诱导的线粒体膜电位破坏在一定程度上得到缓解，同时一氧化氮的产生也有所减少。羧基ptio也有部分缓解作用。我们的研究结果表明，V-ATPase抑制剂巴菲霉素A1和康纳霉素A同样诱导NO的产生，新产生的NO部分参与V-ATPase抑制剂诱导的RAW 264.7细胞凋亡。
In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type (H+)-ATPase (V-ATPase) inhibitors bafilomycin A1 and concanamycin A induced nitric oxide (NO) production through the expression of inducible nitric-oxide synthase mRNA and its protein and decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Bafilomycin A1 and concanamycin A activated nuclear factor (NF)-kappa B and activator protein-1 and decreased the level of I kappa B-alpha and increased that of phosphorylated c-Jun N-terminal kinase (JNK). NO production induced by these V-ATPase inhibitors was suppressed by the NF-kappa B inhibitor Bay 11-7082 [(E)3-[(4-methylphenyl)sulfonyl])2- propenenitrile] and the JNK inhibitor SP600125 [anthra[1,9cd] pyrazol-6(2H)-one] in parallel with the partial alleviation of the V-ATPase inhibitor-induced decrease in MTT response. The Na+, K+-ATPase inhibitor dibucaine and the F-ATPase inhibitor oligomycin did not induce NO production at which concentrations the MTT response was decreased. The NO donor S-nitrosoN- acetyl-DL-penicillamine further lowered the V-ATPase inhibitor- induced decrease in the MTT response, and the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline1- oxyl-3-oxide, sodium salt (carboxy-PTIO) alleviated it partially. Mitochondrial depolarization, an index of apoptosis, was induced by bafilomycin A1 and concanamycin A. On treatment with the nitric-oxide synthase inhibitor N-G-monomethyl-L- arginine acetate, the disruption of mitochondrial membrane potential induced by bafilomycin A1 and concanamycin A was alleviated partially in parallel with the decrease in NO production. Carboxy-PTIO also alleviated it partially. Our findings suggest that the V-ATPase inhibitors bafilomycin A1 and concanamycin A similarly induce NO production and the newly produced NO participates partially in the V-ATPase inhibitor-induced apoptosis in RAW 264.7 cells.