Nitric oxide inhibits bafilomycin-sensitive H(+)-ATPase activity in rat cortical collecting duct.
Nitric oxide inhibits bafilomycin-sensitive H(+)-ATPase activity in rat cortical collecting duct.
复制标题
一氧化氮抑制大鼠皮质集合管中巴弗洛霉素敏感的 H()-ATP 酶活性。
DOI：
10.1152/ajprenal.1994.267.4.f509
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Madsen,KM
中科院分区：
文献类型：
--
作者：
Tojo,A;Guzman,NJ;Garg,LC;Tisher,CC;Madsen,KM
一氧化氮（NO）是一氧化氮合酶（NOS）从L-精氨酸产生的信使分子。一些NOS同工型组成性地存在于细胞中，而另一些可以由细胞因子诱导。最近的证据表明，NO抑制细胞内pH调节的空泡H（+）-腺苷三磷酸酶（ATP酶）在巨噬细胞，其中含有一种诱导型的NOS。液泡H（+）-ATP酶参与集合管闰细胞的质子分泌。因此，我们用荧光微量分析法研究了NO对正常大鼠经胶原酶处理的肾脏中单个皮质集合管（CCD）中巴夫霉素敏感的H（+）-ATP酶活性的影响。将CCD与NO供体硝普钠（0.1和1 mM）或3-吗啉代-悉尼酮亚胺盐酸盐（SIN-1，30 μ M）孵育，导致H（+）-ATP酶活性的剂量依赖性降低。将CCD与脂多糖（LPS）和γ-干扰素（可诱导巨噬细胞中的NOS）一起孵育，可使H（+）-ATP酶活性降低85%。这种作用可通过与NOS竞争性抑制剂N ω-硝基-L-精氨酸同时孵育来阻止，表明H（+）-ATP酶活性的降低是由NO的产生引起的。与8-溴-环磷酸鸟苷（cGMP）共同孵育也抑制H（+）-ATP酶活性，提示NO可能通过激活鸟苷酸环化酶和产生cGMP而在CCD中发挥作用。免疫组化使用抗体的巨噬细胞型NOS显示强烈的标记的嵌入细胞中的CCD，确认在这些细胞中的NOS的存在。（250字处删节）
Nitric oxide (NO) is a messenger molecule that is produced from L-arginine by NO synthase (NOS). Some NOS isoforms are present in cells constitutively, whereas others can be induced by cytokines. Recent evidence suggests that NO inhibits intracellular pH regulation by the vacuolar H(+)-adenosinetriphosphatase (ATPase) in macrophages, which contain an inducible form of NOS. The vacuolar H(+)-ATPase is involved in proton secretion in intercalated cells in the collecting duct. We have therefore examined the effect of NO on bafilomycin-sensitive H(+)-ATPase activity in individual cortical collecting ducts (CCD) microdissected from collagenase-treated kidneys of normal rats using a fluorometric microassay. Incubation of CCD with the NO donors, sodium nitroprusside (0.1 and 1 mM) or 3-morpholino-sydnonimine hydrochloride (SIN-1, 30 microM), caused a dose-dependent decrease in H(+)-ATPase activity. Incubation of CCD with lipopolysaccharide (LPS) and interferon-gamma, which induces NOS in macrophages, decreased H(+)-ATPase activity by 85%. This effect was prevented by simultaneous incubation with N omega-nitro-L-arginine, a competitive inhibitor of NOS, indicating that the decrease in H(+)-ATPase activity was caused by NO production. Incubation with 8-bromo-guanosine 3',5'-cyclic monophosphate (cGMP) also inhibited H(+)-ATPase activity, suggesting that NO may exert its effect in the CCD via activation of guanylyl cyclase and production of cGMP. Immunohistochemistry using antibodies to the macrophage-type NOS revealed strong labeling of intercalated cells in the CCD, confirming the presence of NOS in these cells.(ABSTRACT TRUNCATED AT 250 WORDS)