• Journal of Ginseng Research
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• 제47권2호
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• pp.199-209
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• 2023

Background: Due to the interrupted blood supply in cerebral ischemic stroke (CIS), ischemic and hypoxia results in neuronal depolarization, insufficient NAD+, excessive levels of ROS, mitochondrial damages, and energy metabolism disorders, which triggers the ischemic cascades. Currently, improvement of mitochondrial functions and energy metabolism is as a vital therapeutic target and clinical strategy. Hence, it is greatly crucial to look for neuroprotective natural agents with mitochondria protection actions and explore the mediated targets for treating CIS. In the previous study, notoginseng leaf triterpenes (PNGL) from Panax notoginseng stems and leaves was demonstrated to have neuroprotective effects against cerebral ischemia/reperfusion injury. However, the potential mechanisms have been not completely elaborate. Methods: The model of middle cerebral artery occlusion and reperfusion (MCAO/R) was adopted to verify the neuroprotective effects and potential pharmacology mechanisms of PNGL in vivo. Antioxidant markers were evaluated by kit detection. Mitochondrial function was evaluated by ATP content measurement, ATPase, NAD and NADH kits. And the transmission electron microscopy (TEM) and pathological staining (H&E and Nissl) were used to detect cerebral morphological changes and mitochondrial structural damages. Western blotting, ELISA and immunofluorescence assay were utilized to explore the mitochondrial protection effects and its related mechanisms in vivo. Results: In vivo, treatment with PNGL markedly reduced excessive oxidative stress, inhibited mitochondrial injury, alleviated energy metabolism dysfunction, decreased neuronal loss and apoptosis, and thus notedly raised neuronal survival under ischemia and hypoxia. Meanwhile, PNGL significantly increased the expression of nicotinamide phosphoribosyltransferase (NAMPT) in the ischemic regions, and regulated its related downstream SIRT1/2/3-MnSOD/PGC-1α pathways. Conclusion: The study finds that the mitochondrial protective effects of PNGL are associated with the NAMPT-SIRT1/2/3-MnSOD/PGC-1α signal pathways. PNGL, as a novel candidate drug, has great application prospects for preventing and treating ischemic stroke.

• Biomolecules & Therapeutics
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• 제17권3호
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• pp.270-275
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• 2009

Polyethylene glycol-conjugated hemoglobin (PEG-Hb) has been proposed as a blood substitute for transfusion due to their plasma expansion and oxygen transport capabilities. The protective effect of PEG-Hb on cerebral hypoxic-ischemic injury was investigated in neonatal hypoxia model and adult rat focal cerebral ischemia model. As intravenously administered 30 min before the onset of hypoxia, PEG-Hb markedly protected cerebral hypoxic injury in a neonatal rat hypoxia model. A similar treatment of PEG-Hb largely reduced the ischemic injury ensuing after 2-h middle cerebral artery occlusion followed by 22-h reperfusion. Consistently, neurological disorder was significantly improved by PEG-Hb. The results indicate that the pharmacological blockade of cerebral ischemic injury by using PEG-Hb may provide a useful strategy for the treatment of cerebral stroke.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.647-651
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• 2002

An LC/MS/MS method for the simultaneous determination of a neuroprotective agent for ischemia-reperfusion damage, KR-31378 and its N-acetyl metabolite KR-31612 in human plasma was developed. KR-31378, KR-31612 and the internal standard. KR-31543 were extracted from human plasma by liquid-liquid extraction. A reverse-phase HPLC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-5 mM ammonium formate (55:45, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring mode. The lower limits of quantification for KR-31378 and KR-31612 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity.

• The Korean Journal of Physiology and Pharmacology
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• 제10권6호
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• pp.337-341
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• 2006

The present study was designed to investigate the protein expression of nitric oxide synthase (NOS) and guanylyl cyclase (GC) activity in ischemia/perfusion (I/R) renal injury in rats. Renal I/R injury was experimentally induced by clamping the both renal pedicle for 40 min in Sprague-Dawley male rats. The renal expression of NOS isoforms was determined by Western blot analysis, and the activity of guanylyl cyclase was determined by the amount of guanosine 3', 5'-cyclic monophosphate (cGMP) formed in response to sodium nitroprusside (SNP), NO donor. I/R injury resulted in renal failure associated with decreased urine osmolality. The expression of inducible NOS (iNOS) was increased in I/R injury rats compared with controls, while endothelial NOS (eNOS) and neuronal NOS (nNOS) expression was decreased. The urinary excretion of NO metabolites was decreased in I/R injury rats. The cGMP production provoked by SNP was decreased in the papilla, but not in glomerulus. These results indicate an altered regulation of NOS expression and guanylyl cyclase activity in I/R-induced nephropathy.

• 동의생리병리학회지
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• 제20권1호
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• pp.25-30
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• 2006

This Study was designed to investigate the effects of Patholobi Caulis on the change of regional cerebral blood flow (rCBF) and blood Pressure (MABP) in normal and Cerebral ischemic rats. And, this Study was designed to investigate the inhibition of lactate dehydrogenase (LDH) activity in neuronal cells. The results were as follows : In normal rats, Patholobi Caulis significantly increased rCBF in a dose-dependent manner, and MABP was somewhat increased. In ischemia rats, rCBF was significantly and stably increased by Patholobi Caulis (10 mg/kg, i.p.) during the period of cerebral reperfusion, which contrasted with the findings of rapid and marked increase in control group. Patholobi Caulis significantly inhibited LDH activity in neuronal cells. It was suggested that Patholobi Caulis had an anti-ischemic effect through the improvement of cerebral hemodynamics and inhibitive effect on the brain damage.

• 동의생리병리학회지
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• 제18권6호
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• pp.1643-1651
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• 2004

This experimental study was designed to investigate the effects of SIEGESBECKIAE HERBA extract (SHE) on the change of cerebral hemodynamics 〔regional cerebral blood flow (rCBF) and mean arterial blood pressure(MABP)〕 in normal condition and cerebral ischemic rats, and to determine the mechanism of action of SHE. This study was designed to investigate whether or not SHE inhibit lactate dehydrogenase (LDH) activity in neuronal cells and cytokines production in serum of cerebral ischemic rats. The results were as follows SHE increased rCBF significantly in a dose-dependent manner, but MABP was not changed by SHE in normal rats. The SHE-induced increase in rCBF was significantly inhibited by pretreatment with indomethacin (IDN), an inhibitor of cyclooxygenase but was increased by methylene blue (MTB), an inhibitor of guanylate cyclase. SHE inhibited lactate dehydrogenase (LDH) activity significantly in neuronal cells. rCBF was increased significantly and stably by SHE(10㎎/㎏, i.p.) during the period of cerebral reperfusion, which contrasted with the findings of rapid and marked increase in control group in ischemic rats. In serum by drawing from femoral arterial blood after middle cerebral arterial occlusion(MCAO) for 1hr and reperfusion for 1hr, the sample group was decreased IL-1β production significantly compared to that of the control group. In serum by drawing from femoral arterial blood after MCAO 1hr and reperfusion 1hr, sample group decreased TNF-α production significantly compared to that of the control grolilp. In serum by drawing from femoral arterial blood after reperfusion 1hr, sample group increased TGF-β production significantly compared to that of the control group. In serum by drawing from femoral arterial blood after MCAO for 1hr and reperfusion for 1hr, IL-10 production of the sample group was similar to that of control group. These results suggested that SHE had inhibitive effect on the brain damage by inhibited LDH activity, IL-1β and TNF-α production, but accelerated TGF-β production.

• 대한핵의학회지
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• 제30권4호
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• pp.484-492
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• 1996

허혈성 뇌 병변에서 $^{99m}Tc$-glucarate의 섭취에 관한 연구를 하기 위하여 중뇌동맥 폐쇄 쥐 뇌허혈 모델을 재관류한 군과 하지 않은 군으로 나누어 만들었다. $^{99m}Tc$-glucarate와 [$^{18}F$]FDG를 연속적으로 투여하여 그 분포 양상을 이중 자가방사촬영법으로 관찰하였다. 조직의 괴사 여부를 알기 위하여 TTC 염색도 실시하여 동일한 뇌표본에 대하여 3가지의 영상을 동시에 얻을 수가 있었다. 얻은 영상으로 섭취 또는 비섭취정도를 보아 0점에서 3점까지 점수를 매겨 합하여 비교하였다. 수술한 쥐들 18마리 중 10마리가 신경학적 증상을 보이면서 살아남아 실험대상이 되었다. TTC 염색으로 확인한 경색크기는 재관류하지 않은 군이 컸다. [$^{18}F$]FDG 섭취양상은 TTC 염색과 거의 비슷하였다. 다만 일부에서 TTC 염색되는 곳에 [$^{18}F$]FDG 가 중간정도로 섭취되는 곳이 있었고 TTC 염색되지 않는 곳에 [$^{18}F$]FDG가 중간정도 섭취되는 곳이 있었다. TTC로 염색된 부위에는 $^{99m}Tc$-glucarate가 섭취되지 않았다. TTC로 염색되지 않는 곳에 일부분 $^{99m}Tc$-glucarate가 섭취되었다. TTC와 [$^{18}F$]FDG가 염색되거나 섭취되지 않는 곳에 $^{99m}Tc$-glucarate가 섭취되지 않는 곳이 있었다. 그러나 [$^{18}F$]FDG의 중간 정도의 섭취나 [$^{18}F$]FDG와 TTC사이에 부합하지 않는 곳 등과 $^{99m}Tc$-glucarate 섭취와 대응시키기 어려웠다 $^{99m}Tc$-glucarate가 재관류 군에서 더 넓고 많이 섭취되었다. 결론적으로 $^{99m}Tc$-glucarate는 비생존 허혈조직에만 섭취되는데 관류재개통에 따라 다양하게 섭취되었고 재관류모델에 더 많이 넓게 섭취되었다. 중뇌동맥폐색 및 재관류 모델로 $^{99m}Tc,\;^{18}F$ 및 TTC 동시영상을 얻어 생존능과 포도당대사의 부합 비부합 여부를 밝히고 그 의의를 조사한 후 $^{99m}Tc$-glucarate섭취와 상관을 조사하면 $^{99m}Tc$-glucarate섭취의 의의를 밝힐 수 있을 것으로 본다.

• 대한본초학회지
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• 제21권2호
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• pp.151-158
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• 2006

Objectives : The purpose of the present study is to observe the neuroprotective effect of the $NeuBo153^{\circledR}$ on transient focal cerebral ischemia in rats. Methods : $NeuBo153^{\circledR}$ was made by mixing the herbs, mainly the root of Panax ginseng, the root of Rehmannia glutinosa and Poria cocos, the stem bark of Acanthopanax senticosus, the root of Scutellaria baicalensis and Mel, and heating for 96 hours. Transient Focal cerebral ischemia (2 h of ischemia, 22 h of reperfusion) was induced by intraluminal suture method with SD rats. Sensory motor function was tested by rotarod test, prehensile traction test, beam balance test and foot fault test at 24 h after ischemia. The brain slices were stained by 2% 2, 3, 5-triphenyltetrazolium chloride and the infarct volume was measured by graphic analyzer at 24 h after ischemia. Results : $NeuBo153^{\circledR}$ treated group did not show significant differences compared with vehicle treated group in body temperature. Oral administration of $NeuBo153^{\circledR}$ reduced brain infarct volume by 29.7% compared with vehicle treated group. $NeuBo153^{\circledR}$ also showed protective effects on sensory motor functional deficits. Conclusion : $NeuBo153^{\circledR}$ treatment reduced brain damage and improved functional deficits induced by MCAo. It showed neuroprotective effects even when treatment was relayed 2 h after injury. Further research is required to evaluating long term functional recovery am accurate therapeutic range and mechanisms.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.697-703
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• 2002

We investigated that the role of nitric oxide (NO) on ischemic rats in brain and heart. Ischemia was induced by both common carotid arteries (CCA) occlusion for 24h following reperfusion. Then tissue samples were removed and measured NOx. In brain, NOx was increased by about 40% vs. normal and it was significantly inhibited by aminoguanidine, selective iNOS inhibitor. This result showed that NOx concentration was increased by iNOS. We investigated the role of $Ca^{2+}$ during ischemia. Nimodipine, L-type calcium channel blocker, didn't inhibit the increases of NOx concentration during ischemia. It suggested that increased NOx was due to calcium-independent NOS. MK-801, which N-methyl-D-aspartate (NMDA) receptor antagonist, didn't significantly prevent the increases of NOx. In heart, ischemia caused NOx decrease and it is inconsistent with NOx increase in brain. Aminoguanidine and nimodipine didnt affect on NOx decrease. But MK-801 more lowered NOx concentration than those of ischemia control group. It seemed that $Ca^{2+}$ influx in heart partially occurred via NMDA receptor and inhibited by NMDA receptor antagonist. The mean arterial pressure (MAP) in ischemic rats after 24h of CCA occlusion was decreased when compared to normal value, whereas the heart rates (HR) was not different between two groups. Aminoguanidine or MK801 had no effect on MAP or HR, but nimodipine reduced MAP. There was no difference the effects of aminoguanidine, nimodipine, or MK-801, on MAP and HR between normal rats and ischemic rats. In summary, ischemic model caused an increase of NOx concentration, suggesting that this may be produced via iNOS, which is calcium independent in brain. However in heart, ischemia decreased NOx concentration and NMDA receptor was partially involved. The basal MAP was decreased in ischemic rats but HR was not different from normal control, suggesting that increased NOx in brain of ischemic rat may result in the hypotension.

• 동의생리학회지
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• 제14권2호통권20호
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• pp.179-187
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• 1999

To investigate how Jagamchotang provent cellular injury by a certain starting point on reperfusion injury after ischemia in myocardial cell, conducted MTT assay, LM stydy and measured LDH secretion, heart rate and nitric oxide(NO), and got the following results. 1. Jagamchotang did not injure cells even in $20{\mu}g/ml$. 2. Jaganchotang repressed the toxicity of mitochondria and cell membrane in reperfusing after ischemia and repressed the contraction of promontory of myocardial cell and reduction of the number of cells. Also maintained regular heart rate and reduced the number of heart rate. 3. Synthesis of NO by Jagamchotang in ischemia increased 1.9 times than a control. 4. When reperfusing with sodium nitropruside (SNO), NO donor in ischemia repressed the toxicity of mitochondria as the case of reperfusing with Jagamchotang in ischemia. Therefore, putting these findings together, it. can be said the effect of Jagamchotang in ischemia will be closely related with generation of NO.