• 생물정신의학
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• 제3권2호
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• pp.191-202
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• 1996

This study was aimed to charily the change of NADPH-diaphorase(NADPH-d) and neuropeptide Y(NPY) associated with aging of the rots. To verily the effect of aging of NPY and NADPH-d-positive neurons in the brain stem, the neurons were stained by the immunohistochemical and histochemical method. In the aged group, the number of NADPH-d-positive neurons was significantly decreased in substantia nigra lateralis, supragenual nucleus, raphe magnus nucleus and raphe obscurus nucleus as compared with control group. The number of NADPH-d-positive neurons was not significantly decreased in superior colliculus, interpeduncular nucleus, central gray. dorsal raphe nucleus, retrorubral nucleus, pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus, pontine reticular nucleus, prepositus hypoglossal nucleus and nucleus solitarius of the aged rats. The NADPH-d and NPY-positive neurons were found in the interpeduncular nucleus, central gray, substantia nigra lateralis, laterodorsal tegmental nucleus, nucleus solitarius, raphe magnus nucleus, raphe obscurus nucleus of the control and aged groups. The coexistence of NADPH-d and NPY in the some cell was not found in the brain stem of both groups.

• BMB Reports
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• 제28권6호
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• pp.533-537
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• 1995

The detection with lucigenin under physiological conditions is selective for ${\cdot} O_{2}^{-}$, for it can be accepted that lucigenin indicates actual intramembranal $\cdot O_{2}^{-}- formation$. Lucigenin chemiluminescence (CL) was elicited from the plasma membrane (PM) only by addition of reduced pyridine nucleotide. NADPH was preferred to NADH in PM and hepatocytes. This specificity was masked by $NAD(P)^+$ inhibition. The half maximum rate of CL increase was obtained with 1.5 ${\mu}m$ NADH or 55 ${\mu}m$ NADPH in hepatocytes and 6 ${\mu}m$ NADH or 30 ${\mu}m$ NADPH in plasma membranes. Measurement of these NADPH values required the presence of a NADPH-regenerating system. With NADPH the maximal rate obtained was 10 fold higher than with NADH. NADPH and NADH could produce CL when having access from either side of the membrane. They seemed to react with the identical acceptor because NADH-induced CL was also inhibited by $NADP^+$. The characteristics of ${\cdot}O_{2}^{-}-formation$ produced by pyridine nucleotide will be discussed.

• Applied Biological Chemistry
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• 제11권
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• pp.29-33
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• 1969

1. $NADPH_2$산화효소존재하(酸化酵素存在下)에 있어서 Quinone에 의(依)한 $NADPH_2$의 산화(酸化)를 시험관내(試驗管內)에서 조사(調査)하였다. 그 결과(結果)를 $3{\times}10^{-5}M$.의 각종(各種) Quinone 류(類)로 $^{14}CO_2$고정(固定) 10분전급고정시험중(分前及固定試驗中) 처리(處理)한 Chlorella의 $^{11}CO_2$고정율(固定率)과 비교(比較)하였다. 2. 각종(各種) Quinone 류(類)의 $NADPH_2$ 산화속도(酸化速度)와 $^{14}CO_2$ 고정율(固定率)사이에는 밀접(密接)한 관계(關係)가 있음이 밝혀졌다. $NADPH_2$ 산화(酸化)와 $^{14}CO_2$ 고정(固定)에 대(對)한 Quinone의 영향(影響)은 그 세기가 다음 순서(順序)와 같다. Dichlone>06-K>NQ>BQ 3. Chlorella에 대(對)한 Quinone의 독성(毒性)은 NADPH를 식물체(植物體)에서 없앰으로서 $^{14}CO_2$ 고정(固定)을 방해(妨害)하여 세포(細胞)를 결과적(結果的)으로 죽이는 것이라고 생각이된다. 4. Chlorella의 아미노산(酸) 생합성(生合成)에 대(對)한 Quinone류(類)의 영향(影響)은 전반적(全般的)으로 이를 억제(抑制)하는 것이라 하겠다. 이것은 특(特)히 Dichlone에 있어서 현저(顯著)하다. 이 현상(現象)은 식물(植物)에 $NADPH_2$가 없어지고 $^{14}CO_2$고정(固定)이 조해(阻害)되기 때문일 것이다. 자당(蔗糖)의 생합성(生合成)은 영향(影響)을 받지않거나 오히려 자극(刺戟)을 받는데 그 이유(理由)는 불분명(不分明)하다.

• 약학회지
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• 제56권2호
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• pp.80-85
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• 2012

Previously, we have reported that arachidonic acid (AA) appears to be involved in the induction of apoptosis in HepG2 human hepatoblastoma cells. In this study we investigated the possible role of the NADPH oxidase, a membranebound enzyme generating reactive oxygen species (ROS), in the mechanism of AA-induced apoptosis in HepG2 cells. Apoptotic cell death induced by AA was significantly suppressed by various inhibitors of the NADPH oxidase, diphenylene iodonium (DPI), apocynin (Apo) and neopterine (NP). In addition, these inhibitors of the NADPH oxidase completely blunted the AA-induced ROS elevation. Next, we investigated the implication of metabolic pathway of AA in these AA actions. Both apoptosis and ROS production induced by AA were not significantly altered by treatment with indomethacin (Indo) or nordihydroguaiaretic acid (NDGA), selective inhibitors of cyclooxygenase (COX) and lipoxygenase (LOX), respectively, suggesting that AA metabolites produced by COX or LOX may not have an essential role in the AA-induced apoptosis and ROS generation. Collectively, these results suggest that the NADPH oxidase may be a key player in the mechanism of AA-induced apoptosis in HepG2 cells. These results further suggest that NADPH oxidase may be a good target for the management of human hepatomas.

• Journal of Acupuncture Research
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• 제25권3호
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• pp.179-187
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• 2008

Objectives & Methods : This study was to investigate effect of low frequency electroacupuncture on NADPH-d positive neurons in the brain cortex of rat with adjuvant induced rheumatoid arthritis. Experimental groups were divided into 6 groups ; Normal, Control, $ST_{36}$, $SP_9$, $ST_{36}+SP_9$ and Non-Acupoint. Normal group, non-arthritic group, was injected normal saline, and the other groups were injected FCA. Each acupoint groups were treated by 2Hz electroacupuncture at each acupoints and NA group was treated by 2Hz electroacupuncture at non-acupoint. Each groups were evaluated by the number of NADPH-d positive neurons in primary somatosensory area(S1), secondary somatosensory area(S2), motor area and caudate putamen by using an image analyzer and a microscope. Results : 1. In S1, the number of NADPH-d positive neuron cells in the $ST_{36}$ group were significantly(p<0.05) increased compared with the control group. 2. In S2, the number of NADPH-d positive neuron cells in all electroacupuncture groups were not significantly changed compared with the control group. 3. In motor area, the number of NADPH-d positive neuron cells in $ST_{36}$ group, $SP_9$ group, NA group were significantly(p<0.05) increased compared with the control group. 4. In Caudate putamen, the number NADPH-d positive neuron cells in all electroacupuncture groups were significantly(p<0.05) decreased compared with the control group. Conclusions : Our result demonstrated that low frequency electroacupuncture on $ST_{36}$ & $SP_9$ normalized expression of NADPH-d positive neurons in the brain cortex of the rheumatoid arthritis model in rats.

• Journal of Microbiology and Biotechnology
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• 제27권10호
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• pp.1867-1876
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• 2017

Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP, glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most S. lividans transformants showed better cell growth and higher concentration of cytosolic NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth (6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1 and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9 times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results suggested that the increased NADPH concentration and various intermediates from the PPP specifically triggered undecylprodigiosin biosynthesis that required many precursors and NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic engineering of PPP routes especially suitable for producing secondary metabolites that need diverse primary precursors and NADPH, which is useful information for metabolic engineering in Streptomyces.

• Toxicological Research
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• 제12권2호
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• pp.155-161
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• 1996

The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.

• Molecules and Cells
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• 제38권4호
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• pp.285-296
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• 2015

Oxidative stress has been linked to the pathogenesis of diabetic nephropathy, the complication of diabetes in the kidney. NADPH oxidases of the Nox family, and in particular the homologue Nox4, are a major source of reactive oxygen species in the diabetic kidney and are critical mediators of redox signaling in glomerular and tubulointerstitial cells exposed to the diabetic milieu. Here, we present an overview of the current knowledge related to the understanding of the role of Nox enzymes in the processes that control mesangial cell, podocyte and tubulointerstitial cell injury induced by hyperglycemia and other predominant factors enhanced in the diabetic milieu, including the renin-angiotensin system and transforming growth factor-${\beta}$. The nature of the upstream modulators of Nox enzymes as well as the downstream targets of the Nox NADPH oxidases implicated in the propagation of the redox processes that alter renal biology in diabetes will be highlighted.

• 대한약리학회지
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• 제23권1호
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• pp.33-44
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• 1987

면역보체가 결합되어 있는 zymosan 또는 열로 응집된 IgG에 의하여 활성화된 호중구에서의 NADPH oxidase 의존적인 $O_{\overline{2}}$의 생성과 탐식작용은 칼슘의 흡수과정과 일치하였다. 활성화된 백혈구의 반응은 세포외 칼슘 농도에 따라 항진되었으며, 이는 칼슘의 킬레이트제인 EGTA나 EDTA에 의하여 유의하게 억제되었다. 활성화된 백혈구로부터 $O_{\overline{2}}$의 생성은 dantrolene과 chlorpromazine에 의하여 억제되었다. 칼슘 길항제인 bepredil, diltiazem, verapamil, nifedipine, nimodipine은 효과적으로 활성화된 백혈구의 칼슘흡수, $O_{\overline{2}}$ 생성 그리고 탐식 작용을 억제하였고 NADPH oxidase 활성도 또한 억제하였다. 그러므로, 칼슘 길항제는 칼슘 유입을 억제하거나 칼슘의 세포내 재분포 및 NADPH oxidase 반응계에 작용하여 활성화된 백혈구에서의 $O_{\overline{2}}$의 생성과 백혈구의 탐식작용을 억제할 것으로 시사되었다.

• Journal of Acupuncture Research
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• 제18권4호
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• pp.116-124
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• 2001

Background and Objetive : The aim of this study was to investigate the effect of various electroacupuncture stimulation on NADPH-diaphorase in cerebral cortex, brain stem, cerebellum of spontaneously hypertensive rats. Materials and Methods : We evaluated the changes of NADPH-d-positive neurons using a histochemical method. The staining intensity of NADPH-d-positive neurons was assessed in a quantitative fashion using a microdensitometrical method based on optical density by means of an image analyzer. Results and Conculsion : The average optical density of NADPH-d-positive neurons of 100 Hz (bipolar square wave 0.2 ms duration and 100 Hz frequency) electroacupuncture treatment group significantly increased in most cortical areas comparison between the manual acupuncture and 2 Hz (bipolar square wave 0.2 ms duration and 2 Hz frequency) electroacupuncture groups. In the brain stem, the optical density of NADPH-d-positive neuron at only superficial gray layer of the superior colliculus area was same as cerebral cortex. We conclude that the morphological evidence for NADPH-d-positive neurons may be have regional change in cerebral cortex brain stem and cerebellum according to various electroacupuncture stimulations.