• Journal of Acupuncture Research
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• 제20권6호
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• pp.148-159
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• 2003

Objective: This study examines the effects of acupoint combination on NADPH-diaphorase and neuronal nitric oxide synthase(nNOS) in the brainstem of spontaneously hypertensive rats. Methods: The changes of NADPH-d-positive neurons using a histochemical method and the changes of nNOS-positive neurons using an immunohistochemical method were evaluated. The optical densities of NADPH-d-positive neurons and nNOS-positive neurons of the Choksamni(ST36) Umnungchon(SP9) groups were significantly increased in all brainstem areas as compared to the Choksamni and Choksamni Kokchi(LI11) groups and decreased, with the exeption of the nNOS-positive neurons in the superficial gray of superior colliculus, as compared to the normal group. Results: Our results demonstrated that electroacupuncture changes the activity in the NO system in the brainstem of SHR and the acupoint combination is one of the important parameters for this effect.

• Journal of Nutrition and Health
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• 제33권5호
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• pp.491-496
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• 2000

Food intake is regulated by both central and peripheral mechanisms. In the central nervous, the hypothalamus acts for autonomic and endocrine homeostasis. The paraventricular nucleus(PVN) of hypothalamus is an imprtant site of interaction in central feeding pathways. Neuroepetide Y(NPY)is one of the most powerful neurochemical stimulants of food intake known. Also brain nitric oxide(NO), known as neurotransmitter, is involved in the mechanisms that regulate food intake. In this experiment, 24h fasting mice and anorexia mutant mice have been to examine the expression of NPY, which is the major neuropeptide increasing food intake. Double staining with NPY and nicotinamide-adenine-dinucleotide-phosphate diaphorase(NADPH-d), followed by immunohistochemical method and image analysis, have been used to observe coexisting neurons and the level of expression of each neurons. The results were as follows. 1) NPY-immunoreactivitys reduced immune response of the hypothalamus, particularly paraventricular nucleus(PVN), in anorexia mutant mice. Decreased level of NPY is assumed to be a major pathological factor in anorexia mutant mice. On the other hand, PVN in hypothalamus of fasting mice showed increased immunoreactivity which is in agreement of other researchers. 2) NPY and NADPH-d double staining revealed coexisting neurons in the cerebral cortex. Fasting mice had a tendency to have increased level of coexisting neurons compared to the control group. Compared to the control group, fasting mice express is not increase level of NPY-immunoreactivity, while anorexia mutant mice tended to have a decreased level.

• 동의생리병리학회지
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• 제16권1호
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• pp.192-196
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• 2002

Traditionally, aqueous extracts of Puerariae radix had been used for the treatment of alcohol-related problems. In the present study, the effect of Puerariae radix on cell proliferation and expression of nitric oxide synthase (NOS) in the dentate gyrus of alcohol-intoxicated rats were investigated via 5-bromo-2' -deoxyuridine (BrdU) immunohistochemistry and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry, respectively. Sprague-Dawley rats weighing 150 ± 10 g were divided into four groups: the control group, the Puerarias radix-treated group, the alcohol-treated group, and the alcohol- and Puerariae radix-treated group. The numbers of both BrdU-positive and NADPH-d-positive cells in the dentate gyrus were inhibited significantly by alcohol administration, while Puerariae radix treatment was shown to increase those numbers. In this study, it was revealed that Puerariae radix possesses protective effect against alcohol-induced suppressed new cell formation and NOS expression in the dentate gyrus. Based on the results, it is possible that NO, which might play an important role in the regulation of cell proliferation, is a major target of the toxic effects of alcohol.

• Nutritional Sciences
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• 제8권4호
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• pp.231-236
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• 2005

This study investigated the changes in the neuronal nitric oxide synthase (nNOS) and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activities during food restriction in the rat brain such as cerebral cortex, cerebellum, caudate pautamen and hypothalamus. The rats were placed on a restricted feeding schedule consisting of half the ad libitum quantity for 3 days and 1, 2, 4, 6 and 9 weeks, and a free feeding schedule for 4 weeks. The loss of body weight peaked after 1 week of food restriction and persisted during the entire 9-week period of food restriction. The dramatic weight change in the first week ($12\%$) and the reduction in weight changes thereafter suggest that major adaptation changes occur early and body maintenance occurs subsequently. In the hypothalamus, the optical densities of the NADPH-d and nNOS immunoreactivities were found to be significantly higher in the 1-week and lower in the 9-week food restricted group than in the ad libitum fed control rats. In contrast, in the cerebral cortex, the optical densities of the NADPH-d- and nNOS-positive neurons were not changed significantly during the period of food restriction. This study provides the morphological evidence showing that food restriction has a significant effect on the nitric oxide synthesizing system of the hypothalamus.

• Tuberculosis and Respiratory Diseases
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• 제53권6호
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• pp.595-611
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• 2002

Akt/PKB protein kinase B, ALI acute lung injury, ARDS acute respiratory distress syndrome, CREB C-AMP response element binding protein, ERK extracelluar signal-related kinase, fMLP fMet-Leu-Phe, G-CSF granulocyte colony-stimulating factor, IL interleukin, ILK integrin-linked kinase, JNK Jun N-terminal kinase, LPS lipopolysaccharide, MAP mitogen-activated protein, MEK MAP/ERK kinase, MIP-2 macrophage inflammatory protein-2, MMP matrix metalloproteinase, MPO myeloperoxidase, NADPH nicotinamide adenine dinucleotide phosphate, NE neutrophil elastase, NF-kB nuclear factor-kappa B, NOS nitric oxide synthase, p38 MAPK p38 mitogen activated protein kinase, PAF platelet activating factor, PAKs P21-activated kinases, PMN polymorphonuclear leukocytes, PI3-K phosphatidylinositol 3-kinase, PyK proline-rich tyrosine kinase, ROS reactive oxygen species, TNF-${\alpha}$ tumor necrosis factor-a.

• The Plant Pathology Journal
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• 제21권3호
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• pp.283-287
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• 2005

Since the discovery of generation of $O_2^-$ in plant, many evidence for the oxidative burst (OXB) has been accumulated in various combinations of plant and pathogen or elicitor systems. $O_2^-$ generating system responsible for the OXB was coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in microsomal fraction isolated from sliced aged potato tuber slices which were treated by hyphal wall components elicitor from Phytophthora infestans (HWC). We developed new assay method for quantitative measurement of oxygen radical $O_2^-$ by using electron spin resonance (ESR) analysis during elicitor­induced OXB on the surface of plant tissues. The ESR analysis using an $O_2^-$ trapper, Tiron (1,2-dihydroxy-3,5­benzenedisulfonic acid), provided a convenient assay for detecting only $O_2^-$ during elicitor-induced OXB producing various active oxygen species (AOS) on plant tissue surface. Tiron was oxidized to Tiron semiquinon radical by $O_2^-$. Quantity of the radical signal was measured by specific spectra on ESR spectroscopy. The level of $O_2^-$ was high in from surface of potato tuber tissue treated with hyphal cell wall elicitor (HWC) from Phytophthora infestans. There was no secondary OXB induction by $H_2O_2$ treatment in plant.

• BMB Reports
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• 제38권5호
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• pp.584-590
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• 2005

Glucose 6-phosphate dehydrogenase (EC 1.1.1.49) was purified from Aspergillus aculeatus, a filamentous fungus previously isolated from infected tongue of a patient. The enzyme, apparently homogeneous, had a specific activity of $220\;units\;mg^{-1}$/, a molecular weight of $105,000{\pm}5,000$ Dal by gel filtration and subunit size of $52,000{\pm}1,100$ Dal by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The substrate specificity was extremely strict, with glucose 6-phosphate (G6P) being oxidized by nicotinamide adenine dinucleotide phosphate (NADP) only. At assay pH of 7.5, the enzyme had $K_m$ values of $6\;{\mu}m$ and $75\;{\mu}m$ for NADP and G6P respectively. The $k_{cat}$ was $83\;s^{-1}$. Steady-state kinetics at pH 7.5 produced converging linear Lineweaver-Burk plots as expected for ternary-complex mechanism. The patterns of product and dead-end inhibition suggested that the enzyme can bind NADP and G6P separately to form a binary complex, indicating a random-order mechanism. The enzyme was irreversibly inactivated by heat in a linear fashion, with G6P providing a degree of protection. Phosphoenolpyruvate (PEP), adenosinetriphosphate (ATP), and fructose 6-phosphate (F6P), in decreasing order, are effective inhibitors. Zinc and Cobalt ions were effective inhibitors although cobalt ion was more potent; the two divalent metals were competitive inhibitors with respect to G6P, with $K_i$ values of $6.6\;{\mu}m$ and $4.7\;{\mu}m$ respectively. It is proposed that inhibition by divalent metal ions, at low NADPH /NADP ratio, is another means of controlling pentosephosphate pathway.

• The Korean Journal of Physiology and Pharmacology
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• 제13권5호
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• pp.337-341
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• 2009

Signal transducer and activator of transcription 4 (STAT4), a STAT family member, mediates interleukin 12 (IL12) signal transduction. IL12 is known to be related to calorie-restricted status. In the central nervous system, IL12 also enhances the production of nitric oxide (NO), which regulates food intake. In this study, the expression of neuronal NO synthase (Nos1), which is also related to food intake, was investigated in the hypothalamic areas of Stat4 knockout (KO) mice using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry, a marker for neurons expressing Nos1 enzyme. Western blots were also performed to evaluate Nos1 and Fos expression. Wild-type Balb/c (WT group, n=10 male) and Stat4 KO mice (Stat4 KO group, n=8 male) were used. The body weight and daily food intake in the WT group were $22.4{\pm}0.3$ and 4.4 g per day, while those in the Stat4 KO group were $18.7{\pm}0.4$ and 1.8 g per day, respectively. Stat4 mice had lower body weight and food intake than Balb/c mice. Optical intensities of NADPH-d-positive neurons in the paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of the Stat4 KO group were significantly higher than those of the WT group. Western blotting analysis revealed that the hypothalamic Nos1 and Fos expression of the Stat4 KO group was up-regulated, compared to that in the WT group. These results suggest that Stat4 may be related to the regulation of food intake and expression of Nosl in the hypothalamus.

• Clinical and Experimental Reproductive Medicine
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• 제43권4호
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• pp.193-198
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• 2016

Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect. G6PD plays a key role in the pentose phosphate pathway, which is a major source of nicotinamide adenine dinucleotide phosphate (NADPH). NADPH provides the reducing equivalents for oxidation-reduction reductions involved in protecting against the toxicity of reactive oxygen species such as $H_2O_2$. We hypothesized that G6PD deficiency may reduce the amount of NADPH in sperms, thereby inhibiting the detoxification of $H_2O_2$, which could potentially affect their motility and viability, resulting in an increased susceptibility to infertility. Methods: Semen samples were obtained from four males with G6PD deficiency and eight healthy males as a control. In both groups, motile sperms were isolated from the seminal fluid and incubated with 0, 10, 20, 40, 60, 80, and $120{\mu}M$ concentrations of $H_2O_2$. After 1 hour incubation at $37^{\circ}C$, sperms were evaluated for motility and viability. Results: Incubation of sperms with 10 and $20{\mu}M\;H_2O_2$ led to very little decrease in motility and viability, but motility decreased notably in both groups in 40, 60, and $80{\mu}M\;H_2O_2$, and viability decreased in both groups in 40, 60, 80, and $120{\mu}M\;H_2O_2$. However, no statistically significant differences were found between the G6PD-deficient group and controls. Conclusion: G6PD deficiency does not increase the susceptibility of sperm to oxidative stress induced by $H_2O_2$, and the reducing equivalents necessary for protection against $H_2O_2$ are most likely produced by other pathways. Therefore, G6PD deficiency cannot be considered as major risk factor for male infertility.

• KSBB Journal
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• 제15권2호
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• pp.150-155
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• 2000

본 연구는 Streptomyces sp. BH-405의 배양액으로부터 유기용매로 추출하여 지질과 산화에 대한 항산화 효과를 실험하였다. Streptomyces sp. BH-405의 배양액을 ethylether로 추출한 후 silica gel column(100$\times$10 cm) chromatography로 분획한 획분을 POV로 측정한 결과 fraction 3에서 향산화 활성이 가장 높았다. 항산화 활성이 높은 fraction 3 획분을 다시 preparative TLC와 column chromatography로 더욱 정제하여 band 2를 얻었다. Band 2의 항산화 효과를 알아보기 위해 기존의 항산화제와 비교한 결과 이-$\alpha$-tocopherol, BHA보다 산화억제 효과가 높았으며 DPPH에 의한 수소 공여능은 실험결과가 우수하였다. 한편 band 2는 쥐 간의 microsome에 있어서 유리기의 소거기능이 있었고 NADPH 및 Fenton 시액에 위한 효소적 과산화 지질의 생성을 억제하였다. 그리고 mitochondria와 리놀산의 과산화 유도계에서도 지질의 과산화 과정에 생성하는 유리기의 형성을 차단하였다. 결론적으로 Streptomyces sp. BH-405의 배양액에서 얻은 획분인 band 2는 산화를 억제하는 활성이 있었다.