烟酰胺腺嘌呤二核苷酸及其代谢物在神经细胞死亡中的作用

烟酰胺腺嘌呤二核苷酸(辅酶I)(nicotinamide adenine dinucleotide,NAD+)、还原型烟酰胺腺嘌呤二核苷酸(还原型辅酶Ⅰ)(reduced nicotinamide adenine dinucleotide,NADH)、烟酰胺腺嘌呤二核苷酸磷酸(辅酶Ⅱ)(nicotinamide adenine dinucleotide phosphate,NADP3和还原型烟酰胺腺嘌呤二核苷酸磷酸(还原型辅酶Ⅱ)(reduced nicotinamide adenine dinucIcotide phosphate,NADPH)是能量代谢、维持线粒体功能、抗氧化和还原合成中的关键分子.     

Stimulation of dopamine biosynthesis in cultured PC 12 phaeochromocytoma cells by the coenzyme nicotinamide adeninedinucleotide (NADH)

Summary The activity of the tyrosine hydroxylase, the enzyme which is diminished in the brains of Parkinson patients, has been measured in cultured PC 12 rat phaeochromocytoma cells. In the same way dopamine content in the medium after incubating these cells with or without NADH was assayed. The experiment shows that NADH is able to increase the activity of the tyrosine hydroxylase and dopamine — production in PC 12 cells up to 6 times.The results provide evidence that NADH is able to stimulate dopamine — biosynthesis directly.     

No evidence for cognitive improvement from oral nicotinamide adenine dinucleotide (NADH) in dementia

Summary. Reduced nicotinamide adenine dinucleotide (NADH) is advertised as an over-the-counter product or dietary supplement to treat Alzheimer's disease. We performed a 3-month open-label study with oral 10 mg/day NADH with 25 patients with mild to moderate dementia of the Alzheimer, vascular, and fronto-temporal types in addition to their current cholinomimetic drug medication. In 19 patients who completed the study, we found no evidence for any cognitive effect as defined by established psy-chometric tests. We conclude that NADH is unlikely to achieve cognitive improvements in an extent reported earlier, and present theoretical arguments against an effectiveness of this compound in dementia disorders. Received March 27, 2000; accepted September 20, 2000     

Stimulation of endogenous L-dopa biosynthesis — a new principle for the therapy of Parkinson's disease. The clinical effect of nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotidephosphate (NADPH)

Abstract– The coenzyme nicotinamide adenine dinucleotide (NADH) has been used as a novel medication in 161 Parkinson patients in an open label trial. In all but 18 patients (11.2%) an improvement in their disability was observed. 115 patients (71.4%) showed a very good (better than 30%) response, and 28 patients (17.4%) a moderate response up to 30%. The best results were obtained with a dose of 25 to 50 mg every second day by i.v. administration. Concomitantly with the improvement in disability, the urine HVA level increased significantly, indicating a stimulation of endogenous L-DOPA biosynthesis. 8 patients have been treated with nicotinamide adenine dinucleotidephosphate (NADPH), 5 of whom exhibited an improvement in their disability from 35 to 55%. The other 3 showed a moderate response of 20 to 25%. In all these patients an increase in the urine level of HVA was detected, reflecting elevated endogenous L-DOPA production.     

Treatment with reduced nicotinamide adenine dinucleotide (NADH) improves water maze performance in old Wistar rats

Age-associated cognitive impairment and related neurodegenerative disorders are an increasing major public health problem. Nicotinamide adenine dinucleotide (NADH), a co-substrate for energy transfer in the mitochondrial respiratory chain, is speculated to induce positive effects in some of these diseases. Studies showed diminished mitochondrial function in patients with M. Alzheimer. In a preliminary clinical trial NADH given peripherally improved cognitive function in Alzheimer disease. Previous own experiments revealed an increased NADH level in the rat brain following peripheral application of NADH (10-100 mg/kg, i.p.+ i.v.). Therefore, we wanted to know, whether or not NADH has an effect on cognitive function in animals. We analysed the effect of repeated i.p. injection of NADH on the performance of 3-month-old and 22-month-old Wistar rats in the Morris water maze and in the rota-rod test of motor coordination. The rats were injected for 10 days once daily with the doses of NADH used in the bioavailability study (10-100 mg/kg) or vehicle 20 min before the behavioural tests. The repeated administration of NADH improved the performance of old rats in the acquisition phase (place version) and the spatial probe of the Morris water maze compared to vehicle-treated controls. The effect of NADH on learning-related processes is supported by the lack of effects on motor performance on the rota-rod. In summary, our results suggest cognitive enhancing properties of NADH in learning impaired old rats.     

Neurons co-localizing calretinin immunoreactivity and reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the hippocampus and dentate gyrus of the rat

Co-localization of calretinin immunoreactivity and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity was studied in the rat hippocampus and dentate gyrus. Neurons co-expressing both markers (CR/NADPH-d) were observed throughout the hippocampus and dentate gyrus. However, they were more abundant in the stratum pyramidale and radiatum of CA3, stratum pyramidale of CA1, and in the juxtagranular zone of the hilus. The NADPH-d activity appeared in 37% of the calretinin immunoreactive neurons in CA3, 42% in CA1, and 36% in the dentate gyrus, whereas calretinin immunoreactivity occurred in 41% of the NADPH-d positive neurons in the hippocampus, and 16% in the dentate gyrus. The morphology and location of the double marked cells could not be used as a characteristic of the co-localizing neurons. The heavily stained NADPH-d neurons occurring mainly in CA1 do not show calretinin immunoreactivity. NADPH-d fiber swellings could be observed in close apposition to calretinin immunoreactive neurons and dendrites, suggesting synaptic contacts. It has been reported that calretinin immunoreactivity and NADPH-d activity co-localize infrequently in other areas such as the neocortex, striatum, hypothalamus and tegmental nucleus. The relatively high proportion of double marked cells found in the hippocampus and dentate gyrus could be indicative of the importance of the CR/NADPH-d interneurons in the circuitries of these areas.     

Altered nicotinamide adenine dinucleotide (NADH) fluorescence in dt sz mutant hamsters reflects differences in striatal metabolism between severe and mild dystonia

The dt(sz) mutant hamster represents a unique rodent model of idiopathic paroxysmal dystonia. Previous data, collected post-mortem or in anesthetized hamsters under basal conditions, indicated the critical involvement of enhanced striatal neuronal activity. To assess the importance of an enhanced striatal neuronal activity directly during a dystonic episode, continuous monitoring of changes in brain metabolism and therefore neuronal activity indirectly in awake, freely moving animals is necessary. Determination of CNS metabolism by NADH measurement by laser-induced fluorescence spectroscopy in conscious dt(sz) and nondystonic control hamsters revealed reversible decreased NADH fluorescence during dystonic episodes. The degree of change corresponded to the severity of dystonia. This study represents the first application of this innovative method in freely moving animals exhibiting a movement disorder. Our data clearly confirm that the expression of paroxysmal dystonia in dt(sz) mutant hamsters is associated with enhanced striatal neuronal activity and further underscore the versatile application of NADH fluorescence measurements in neuroscience.     

Colocalization of nitric oxide synthase and some neurotransmitters in the intramural ganglia of the guinea pig urinary bladder

The distribution of nitrergic neurons was investigated by using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry in wholemount preparations of the urinary bladder in guinea pigs. Both NADPH-d⁺ and NOS⁺ neurons were located predominantly in the bladder base. Double staining showed that 70.9% of the NADPH-d⁺ neurons coexpressed NOS. Acetylcholinesterase histochemistry revealed that a majority of the intramural neurons were reactive, and about half of them (51.4%) were double labelled for NOS. Tyrosine hydroxylase-positive neurons were also distributed mainly in the bladder base but in a neuronal population that was separate from the preponderant NADPH-d⁺ neurons. Vasoactive intestinal polypeptide immunoreactivity was also detected in the some of intramural ganglion cells, in which 21.3% of them coexpressed NADPH-d. Calcitonin gene-related peptide and substance P immunoreactivities were confined to nerve fibers, often in close association with NADPH-d⁺ cells or extended along the blood vessels. These results have demonstrated the colocalization of NADPH-d and NOS in the majority of intramural ganglion cells. Many of the nitrergic neurons are apparently cholinergic, indicating that they are parasympathetic postganglionic neurons, and this underscores NO as the major neuromodulator in the parasympathetic nerves in the bladder walls. The localization of vasoactive intestinal polypeptide in nitrergic neurons suggests that the peptide may complement NO for regulation of micturition reflex. The close relationship of NADPH-d-reactive intramural neurons with calcitonin gene-related peptide and substance P fibers, most probably derived from dorsal root ganglion cells, suggests that NO released from the local neurons may exert its influence on the sensory neural pathways in the urinary bladder. J. Comp. Neurol. 394:496–505, 1998. © 1998 Wiley-Liss, Inc.     

Reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry in the hippocampal formation of the New World monkey (Saimiri sciureus)

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) containing fibers and neurons within the hippocampal formation and entorhinal cortex of the new world monkey were determined using a direct histochemical procedure. Occasional intensely stained bipolar NADPH-d positive neurons were seen in the polymorphic zone within the hilus of the dentate gyrus and molecular layer of the hippocampus. Although virtually no intensely stained cells were seen in the CA subfields, a few small oval lightly stained NADPH-d perikarya were found subjacent to CA2. An occasional intensely stained multipolar NADPH-d containing neuron was observed in the subiculum, presubiculum and parasubiculum. In the entorhinal cortex, NADPH-d cells were scattered in all layers with the greatest preponderance in layers 5-6 and underlying white matter. Dense bands of NADPH-d fibers occurred in the outer layer of the molecular layer of the dentate gyrus and the hippocampo-subicular border. NADPH-d fibers also were seen in pre- and parasubicular regions. NADPH-d fiber staining in entorhinal cortex varied mediolaterally with an increasing laminar distribution more caudally. The heaviest bands of NADPH-d fibers occurred in layers 1 and 4 and the white matter-layer 6 border. The distribution patterns of this select neuronal population may be relevant to the study of hippocampal and entorhinal areas in neurodegenerative diseases.     

Glutamine inhibits the accumulation and hydroxylation of tryptophan in rat striatal synaptosomes

l-Glutamine at the concentration present in cerebrospinal fluid decreases the steady-state accumulation of the aromatic amino acids tryptophan, tyrosine and dihydroxyphenylalanine (DOPA) in rat striatal synaptosomes. Glutamine significantly inhibits synaptosomal tryptophan hydroxylase activity; it has less marked effects on tyrosine hydroxylase and DOPA decarboxylase activities. Thus, interaction between glutamine and tryptophan transport into nerve terminals may be one of the factors regulating the rate of serotonin synthesis in vivo.     

Prenatal development of nicotinamide adenine dinucleotide phosphate-diaphorase activity in the human hippocampal formation

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry was used to study the development of the neurons metabolizing nitric oxide in the prenatal human hippocampal formation. Strongly reactive non-pyramidal neurons appeared in small numbers in the subplate at 15 weeks, and rapidly increased in this layer, as well as the cortical plate—derived layers between 17 and 24 weeks. The marginal zone also had a few NADPH-d cells at 15 weeks. The pattern of these darkly reactive cells stabilized by 28 weeks, with the somata distributed mostly at the border of the cortex and white matter in the entorhinal cortex and subiculum, or the alveus in Ammon's horn. Moderately stained non-pyramidal neurons appeared in the dentate gyrus by 17 weeks, and increased in this region and Ammon's horn up to 28 weeks. Small, lightly reactive non-pyramidal neurons were first seen by 32 weeks and increased in number by term. They were mainly distributed in layers II/III of the entorhinal cortex and stratum pyramidale of the subiculum and Ammon's horn. NADPH-d positive fibers in the marginal zone were mostly thin and developed between 20 and 28 weeks. In other cortical layers, thick processes from the darkly stained NADPH-d neurons appeared first, then fine fibers appeared more numerous, especially after 28 weeks. NADPH-d processes that arose from non-pyramidal cells were frequently apposed to blood vessels, including those in the hippocampal fissure. In addition, NADPH-d reactivity was also present in pyramidal and granule cells, but this staining was most pronounced between 15 and 24 weeks. The results show three types of distinctly stained NADPH-d interneurons in the fetal human hippocampal formation with different developmental courses and morphology. Also, hippocampal principal neurons transiently express NADPH-d at early fetal ages. Our data correlated with other findings suggest that nitric oxide may play a role in neuronal development in the hippocampal formation by modulating neuronal differentiation and maturation, and regulating blood supply. Hippocampus 7:215–231, 1997. © 1997 Wiley-Liss, Inc.     

Distribution of reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) cells and fibers in the monkey amygdaloid complex.

The NADPH-d histochemical method stains a selective population of neurons in the central nervous system. Although the functional significance of the enzyme in these cells is unknown, it has nonetheless proved to be a useful marker. In the present study we describe the distribution of NADPH-d-positive cells and fibers in the amygdaloid complex of the Macaca fascicularis monkey. NADPH-d-positive neurons were distributed throughout the amygdaloid complex. Based on the intensity of the reaction product, three different types of NADPH-d-positive cells were described: type 1 cells, the most intensely stained, varied in morphology and were most commonly found in the accessory basal, basal, and lateral nuclei and in the nucleus of the lateral olfactory tract; type 2 cells, the most common NADPH-d-positive cells, were more lightly stained, were generally stellate in shape, and were found in the lateral, basal, and accessory basal nuclei; type 3 cells were very lightly stained, oval or round in shape, and mostly found in the medial, anterior cortical, and paralaminar nuclei. NADPH-d staining was also associated with axonal fiber plexuses in various regions of the amygdala. The highest densities of stained fibers were found in the lateral nucleus, the parvicellular portion of the accessory basal nucleus, and the anterior amygdaloid area. The lowest densities of NADPH-d-positive fiber staining were found in the amygdalohippocampal area, in the lateral part of the central nucleus, and in the intercalated nuclei. In addition to the neuronal and fiber staining, a diffuse, blue neuropil staining was also observed, most commonly in the anterior cortical nucleus, the medial nucleus, the intercalated nuclei, and especially in the amygdalohippocampal area. The distribution of NADPH-d staining often respected nuclear boundaries within the amygdala and was particularly helpful in clarifying the borders of the amygdalohippocampal area.     

NADPH-diaphorase activity increases during estrous phase in the bed nucleus of the accessory olfactory tract in the female rat

We investigated the presence of nitric oxide in the bed nucleus of the accessory olfactory tract (BAOT) in males, diestrous females and estrous females using NADPH-diaphorase. Our results demonstrate a significant increase in the density of the medium-stained cells in the estrous female rats suggesting that during estrous a specific subpopulation of nitrinergic cells are activated in the BAOT. This might be related to the physiological and behavioral changes that occurs in estrous.     

Reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) profiles in the amygdala of human and New World monkey (Saimiri sciureus).

The topographic distribution of nicotinamide adenine dinucleotide-diaphorase (NADPH-d) stained profiles in the amygdala of the human and new world monkey (Saimiri sciureus) were studied histochemically. Fiber and terminal staining were heterogeneously distributed within the amygdala. The most intense staining occurred in the basolateral subdivision, consisting of the lateral, basolateral and accessory basal nuclei. Moderate staining intensity was observed throughout the cortical and media nuclei and cortical transition area, constituents of the corticomedial subdivision. The central amygdaloid area was characterized by minimal NADPH-d histochemical reactivity. NADPH-d positive neurons were pleomorphic and divisible into two classes based on their staining characteristics: intensely or lightly stained neurons. Their distribution was generally complementary, with the majority of intensely stained neurons occupying the basolateral subdivision. There were no appreciable species differences in the patterns of neuronal, fiber and terminal staining between monkey or human amygdala. These results may be relevant to our understanding of the selective vulnerability of neural systems within the human amygdala in neurodegenerative diseases.     

Postnatal development of nicotinamide adenine dinucleotide phosphate-diaphorase-positive neurons in the retina of the golden hamster

The histochemical method was used to investigate the postnatal development of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) -positive neurons in retinas of the golden hamster. NADPH-d-positive neurons were discernible in the retina at postnatal day (P)1. From P4 onward to adulthood, when the retina acquired its laminated characteristics, NADPH-d- positive neurons were observed in the inner nuclear layer (INL) and the ganglion cell layer (GCL). Results showed that NADPH-d-positive neurons in INL and GCL followed different time courses and patterns in their development. NADPH-d-positive neurons in INL underwent a sharp increase from P4 to P8 (3.6-fold), followed by a decrease to 46% of the maximum at P12. This value was maintained relatively constant to the adult level. The mean diameters of NADPH-d-positive neurons in INL, which were smaller than those in the GCL for all ages, increased from P8 to P12 and from P20 to adulthood. As for neurons in the GCL, the increase in cell number was not so apparent for the earlier postnatal days until P20; thereafter, an obvious increase to the adult level was observed. The mean diameters of the NADPH-d-positive cell bodies in the GCL increased with age, except for P16-P20, during which time there was a slight and insignificant decrease. The tendency of changes in cell density was basically similar to that of the total number for both the INL and the GCL. Between P12 and P20, the density distribution map of the NADPH-d-positive neurons underwent dramatic changes: The highest density shifted from the upper central retina at the earlier postnatal days to the lower central retina in the adult. The two waves of increase in NADPH-d-positive neurons coincide with the process of axonal elongation and synaptogenesis and the acquisition of visual function and experience. It is suggested that these NADPH-d-positive neurons are related to these two developmental events.     

Topographical distribution of reduced nicotinamide adenine dinucleotide phosphate-diaphorase in the brain of the Japanese quail

The distribution of reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity was histochemically investigated in the Japanese quail brain. This enzyme is now considered responsible for the synthesis of nitric oxide, a novel neural messenger whose distribution has not been described in the avian brain until now. The histochemical technique provides a simple and reliable method for staining selected populations of neurons throughout the avian brain. In the telencephalon several regions showed heavily stained NADPH-diaphorase positive neurons and processes. In particular the paleostriatal-paraolfactory lobe complex showed the greatest presence of both positive cells and processes. Neurons and processes were also observed in several regions of the hyperstriatum as well as in the archistriatal nucleus taeniae. Some regions, such as the ectostriatum and the hippocampus, had no positive elements. In the diencephalon, the magnocellular hypothalamic system, which in mammals shows NADPH-diaphorase activity, did not show any particular accumulation of reaction product. On the contrary, retinorecipient areas, such as the visual suprachiasmatic nucleus and the lateral geniculate complex, displayed a composite structure of both positive neurons and processes. The brainstem revealed a large NADPH-diaphorase positive population extending through the tegmental nuclei to the locus coeruleus and subcoeruleus. A complex organization was also observed in the optic lobe, where fusiform elements were distributed within the stratum griseum and superficialis of the tectum. In the medulla, a dense terminal field was observed at the level of the nucleus of the solitary tract, whereas scattered neurons were located within the reticular nuclei. Although the staining of neurons and tracts was highly selective, the positive cells did not correspond to any single known neurotransmitter, neuropeptide, or neuroactive molecule system. Several sensory pathways were heavily stained for the NADPH-diaphorase, including part of the olfactory, visual, and auditory pathways. The findings of the present study reveal that the NADPH-diaphorase-containing systems in the avian brain are organized according to a pattern comparable, because of its complexity, to that observed in mammals. However, important interspecific differences suggest that this novel neural system might be involved in diverse tasks. © 1994 Wiley-Liss, Inc.     

NADPH diaphorase staining within the developing olfactory bulbs of normal and unilaterally odor-deprived rats

Littermate rat paps underwent either unilateral surgical occlusion of the right external naris or sham surgery on postnatal day 1. At 10, 20 or 30 days postpartum olfactory bulbs were sectioned and stained using nicotinamide adenine dinucleotide phophate diaphorase (NADPH-d) histochemistry. Two types of staining were observed and analyzed. The reaction produced a Golgi-like filling of short-axon cells in both deep and superficial bulb areas. No differences in the number, morphology or distribution of these cells were found either across ages or treatment conditions, indicating that the cells are resistant to the effects of the deprivation paradigm. Large regional variations in glomerular and olfactory nerve layer staining density were also observed at each age, reinforcing notions of functional or structural differences between glomeruli at very early ages.     

A comparative cluster analysis of nicotinamide adenine dinucleotide phosphate (NADPH)‐diaphorase histochemistry in the brains of amphibians

Nicotinamide adenine dinucleotide phosphate–diaphorase (NADPH‐d) is a key enzyme in the synthesis of the gaseous neurotransmitter nitric oxide. We compare the distribution of NADPH‐d in the brain of four species of hylid frogs. NADPH‐d–positive fibers are present throughout much of the brain, whereas stained cell groups are distributed in well‐defined regions. Whereas most brain areas consistently show positive neurons in all species, in some areas species‐specific differences occur. We analyzed our data and those available for other amphibian species to build a matrix on NADPH‐d brain distribution for a multivariate analysis. Brain dissimilarities were quantified by using the Jaccard index in a hierarchical clustering procedure. The whole brain dendrogram was compared with that of its main subdivisions by applying the Fowlkes–Mallows index for dendrogram similarity, followed by bootstrap replications and a permutation test. Despite the differences in the distribution map of the NADPH‐d system among species, cluster analysis of data from the whole brain and hindbrain faithfully reflected the evolutionary history (framework) of amphibians. Dendrograms from the secondary prosencephalon, diencephalon, mesencephalon, and isthmus showed some deviation from the main scheme. Thus, the present analysis supports the major evolutionary stability of the hindbrain. We provide evidence that the NADPH‐d system in main brain subdivisions should be cautiously approached for comparative purposes because specific adaptations of a single species could occur and may affect the NADPH‐d distribution pattern in a brain subdivision. The minor differences in staining pattern of particular subdivisions apparently do not affect the general patterns of staining across species. J. Comp. Neurol. 522:2980–3003, 2014. © 2014 Wiley Periodicals, Inc.