• 동의생리병리학회지
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• 제19권3호
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• pp.647-655
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• 2005

This study demonstrated anti-oxidative and neuroprotective effects of Rhei Rhizoma. Anti-oxidative effects were studied on BV-2 microglia cells damaged by $H_2O_2$ and nitric oxide. Neuroprotective effects were studied by using oxygen/glucose deprivation of the organotypic hippocampal slice cultures. The results obtained are as follows; The groups treated with 0.5 and 5 mg/ml of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in CA1 region of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The group treated with 50 mg/ml of Puerariae Radix demonstrated decreases of neuronal cell death area and cell death area percentages in CA1 region, but these were not significant statistically. The groups treated with 0.5 and 5 mg/ml of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in dentate gyrus of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The group treated with 50 mg/ml of Puerariae Radix demonstrated decreases of neuronal cell death area and cell death area percentages in dentate gyrus, but these were not significant statistically. The groups treated with 0.5 and 5 mg/ml of Puerariae Radix revealed significant decreases of TUNEL-positive cells in both CA1 region and dentate gyrus of ischemic damaged hippocampus cultures. The group treated with 50 mg/ml of Puerariae Radix demonstrated significant decrease of TUNEL-positive cells in CA1 region, but not in dentate gyrus of ischemic damaged hippocampus. The groups treated with 0.5 and 5 mg/ml of Puerariae Radix revealed significant decreases of LDH concentrations in culture media of ischemic damaged hippocampus cultures. The group treated with 50 mg/ml of Puerariae Radix demonstrated decrease of LDH concentrations in culture media, but it was not significant statistically. The groups treated with 0.5 and 5 mg/ml of Puerariae Radix revealed significant increases of cell viabilities of BV-2 microglia cells damaged by $H_2O_2$. The group treated with 50 mg/ml of Puerariae Radix demonstrated increase of cell viability of BV-2 microglia cells, but it was not significant statistically. The group treated with 0.5 mg/ml of Puerariae Radix revealed significant increase of cell viability of BV-2 microglia cells damaged by nitric oxide. The groups treated with 5 and 50 mg/ml of Puerariae Radix demonstrated increases of cell viabilities of BV-2 microglia cells, but these were not significant statistically. These results suggested that Puerariae Radix revealed neuroprotective effects through the control effect of apoptosis and oxidative damages.

• 대한한방내과학회지
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• 제25권4호
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• pp.227-241
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• 2004

Objective : This study was performed to investigate neuroprotective effects of Bupleuri Radix against oxidative and ischemic damages. Method : To observe the neuroprotective effects against ischemic damage, ischemic insult was induced by oxygen/glucose deprivation (OGD) on organotypic hippocampal slice cultures (OHSC) from 1 week-old Sprague-Dawley rats. Propidium iodide (PI) fluorescence-stained neuronal dead-cell areas, area percentages and TUNEL-positive apoptotic cells in CA1 and dentate gyrus, and LDH levels in culture media of the OHSC were measured following Bupleuri Radix extract treatment. Result : The following results were obtained: (1) The $5\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in CA1 region of the OHSC from 18 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ of Bupleuri Radix treatment was also significant from 6 hrs to 48 hrs following the OGD and was more effective. (2) The 5 and $50\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in DG region of the OHSC from 6 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ treatment was more effective than the $5\;{\mu}g/ml$ treatment. (3) Bupleuri Radix treatment demonstrated a significant decrease in TUNEL-positive apoptotic cells in CA1 region (with 5 and $50\;{\mu}g/ml$) and in DG region (with $50\;{\mu}g/ml$) of the OHSC damaged by the OGD. (4) Bupleuri Radix treatment demonstrated a significant decrease in LDH concentrations in culture media of the OHSC damaged by the OGD. Conclusion : These results suggest that Bupleuri Radix has neuroprotective and control effects on inflammatory and immune responses where there has been ischemic damage to the central nervous system.

• 대한본초학회지
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• 제30권3호
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• pp.49-54
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• 2015

Objectives : This study investigated the neuroprotective effect of Gastodia Rhizoma-gamibang (GG) water extracts against scopolamine-induced neurotoxicity in the hippocampus of male Sprague-Dawley rats. Methods : The animals were divided into five different groups with six animals per each group. The normal group (Nor) was administered with saline, while the control group (Con) was administered with saline after scopolamine treatment. The experimental group (Exp) was administered orally GG extracts at doses of 200 mg/kg (GG1 group), 400 mg/kg (GG2 group), 1000 mg/kg (GG5 group) for 30 day after scopolamine treatment. Results : From a light microscopy study, the nuclei of neurons and glial cells in the hippocampus were more shrunken or condensed in the 30 day control group compared with normal group. In the experimental groups, proportional to the dose, recovered from neurotoxicity induced by scopolamine. The control group, the density of hippocampal neurons were reduced 17-20% compared to normal group. The densities of neurons from the CA1, and CA3 area of the hippocampus in the GG1, GG2 and GG5 groups significantly increased compared with the Con. In the experimental group, neuronal cells are recovered from scopolamine-induced damage. A number of glial cells are observed increase from GG2 and GG5 groups. The PAS-positive materials in the tissues hippocampus), was lower in the Exp than the Con. Conclusions : The present study demonstrates that Gastodia Rhizoma-gamibang extract reduces neuronal damage in the hippocampus of scopolamin-induced impairment mice.

• International Journal of Oral Biology
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• 제49권1호
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• pp.10-17
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• 2024

Glutamate-mediated oxidative stress causes neuronal cell death by increasing intracellular Ca²⁺ uptake, reactive oxidative species (ROS) generation, mitogen-activated protein kinase (MAPK) activation, and translocation of apoptosis-inducing factor (AIF) to the nucleus. In the current study, we demonstrated that corydaline exerts potent neuroprotective effects against glutamate-induced neurotoxicity. Treatment with 5 mmol/L glutamate increased cellular Ca²⁺ influx, ROS generation, MAPK activation, and AIF translocation. In contrast, corydaline treatment decreased cellular Ca²⁺ influx and ROS generation. Western blot analysis revealed that glutamate-mediated MAPK activation was attenuated by corydaline treatment. We further demonstrated that corydaline treatment inhibited the glutamate-mediated translocation of AIF to the nucleus. We propose that corydaline is a promising lead structure for the development of safe and effective neuroprotectants.

• The Korean Journal of Physiology and Pharmacology
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• 제10권1호
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• pp.13-17
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• 2006

Experimentally induced cortical disorganization exhibits many anatomical features which are characteristic of cortical malformations in children with early-onset epilepsy. We used an immunocytochemical technique and extracellular field potential recordings from the dorsal hippocampus to determine whether the excitability of the CA1 pyramidal cells was enhanced in rats with exnerimentallv induced hippocampal dysplasia. Compared with control rats, the MAM-treated rats displayed a decrease of paired pulse inhibition. When $GABA_A$ receptor antagonists were blocked with $10{\mu}M$ bicuculline the amplitude of the second population spike of the MAM-treated of rats was similar to that of the first population spike, as was in the control rats. The MAM-treated rats had fewer somatostatin and parvalbumin-immunoreactive neurons than the control rats. These results suggest that the enhanced neuronal responsiveness of the in vivo recording of the CA1 in this animal model may involve a reduction of CA1 inhibition.

• The Korean Journal of Physiology and Pharmacology
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• 제19권6호
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• pp.515-522
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• 2015

Notch signaling is a key regulator of neuronal fate during embryonic development, but its function in the adult brain is still largely unknown. Mind bomb-2 (Mib2) is an essential positive regulator of the Notch pathway, which acts in the Notch signal-sending cells. Therefore, genetic deletion of Mib2 in the mouse brain might help understand Notch signaling-mediated cell-cell interactions between neurons and their physiological function. Here we show that deletion of Mib2 in the mouse brain results in impaired hippocampal spatial memory and contextual fear memory. Accordingly, we found impaired hippocampal synaptic plasticity in Mib2 knock-out (KO) mice; however, basal synaptic transmission did not change at the Schaffer collateral-CA1 synapses. Using western blot analysis, we found that the level of cleaved Notch1 was lower in Mib2 KO mice than in wild type (WT) littermates after mild foot shock. Taken together, these data suggest that Mib2 plays a critical role in synaptic plasticity and spatial memory through the Notch signaling pathway.

• 생약학회지
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• 제52권3호
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• pp.149-156
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• 2021

Excessive glutamate can cause oxidative stress in neuronal cells and this can be the reason for neurodegenerative disease. In this study, we investigated the protective effect of Spirulina maxima hot ethanol extract on mouse hippocampal HT22 cell of which glutamate receptor has no function. HT22 cells were pre-treated with S. maxima sample at a dose dependent manner (1, 10 and 100 ㎍/ml). After an hour, glutamate was treated. Cell viability, reactive oxygen species (ROS) accumulation, Ca²⁺ influx, decrease of mitochondrial membrane potential level and glutathione related assays were followed by then. S. maxima ethanol extract improved the cell viability by suppressing the ROS and Ca²⁺ formation, retaining the mitochondrial membrane potential level and protecting the activity of the antioxidant enzymes compared with group of vehicle-treated controls. These suggest that S. maxima may decelerate the neurodegeneration by attenuating neuronal damage and oxidative stress.

• 생명과학회지
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• 제30권6호
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• pp.513-521
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• 2020

Bmi1은 PcG 단백의 일종으로 PRC를 구성하는 성분이다. Bmi1에 관한 초기의 연구는 주로 암세포에서의 역할에 집중되었고, 그 결과 Bmi1은 암세포의 증식과 생존에 중요한 역할을 하는 것으로 받아들여지고 있다. 그러나, 최근의 연구 결과 Bmi1은 퇴행성 신경계 질환이 있는 뇌에서 발현이 감소되어 있고 미토콘드리아의 기능과 활성 산소 수준을 조절하는 것으로 알려져 있다. 본 연구에서는 Bmi1 발현 저하 동물을 이용하여 간질에서 Bmi1의 약물 치료 표적 가능성을 밝히고자 하였다. Bmi1의 발현은 pilocarpine에 의한 경련중첩증 이후 해마의 CA1, CA3 그리고 치상회에서 뚜렷하게 증가하였다. 경련 양상을 볼 때 경련중첩증이 유도되는 비율은 Bmi1^+/+와 Bmi1^+/- 생쥐에서 각각 43.14%와 53.57%로 나타났다. 그러나, 치사율과 해마의 신경세포 손상에는 두 군 간에 통계적으로 유의한 차이를 보이지 않았다. 경련중첩증 유도 2개월 후에 나타난 간질 경련의 빈도수는 통계적 유의성은 없었으나 Bmi1^+/- 생쥐에서 약 50% 낮게 나타났다. 반면, 이끼섬유발아는 Bmi1^+/- 생쥐에서 통계적으로 유의하게 증가하였다. 이러한 결과를 종합하면, Bmi1의 발현이 감소할 때 pilocarpine에 의한 경련 유도와 이끼섬유발아가 증가함을 보여준다.

• Journal of Ginseng Research
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• 제43권2호
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• pp.326-334
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• 2019

Background: The objective of our study was to analyze the neuroprotective effects of ginsenoside derivatives Rb1, Rb2, Rc, Rd, Rg1, and Rg3 against glutamate-mediated neurotoxicity in HT22 hippocampal mouse neuron cells. Methods: The neuroprotective effect of ginsenosides were evaluated by measuring cell viability. Protein expressions of mitogen-activated protein kinase (MAPK), Bcl2, Bax, and apoptosis-inducing factor (AIF) were determined by Western blot analysis. The occurrence of apoptotic and death cells was determined by flow cytometry. Cellular level of $Ca^{2+}$ and reactive oxygen species (ROS) levels were evaluated by image analysis using the fluorescent probes Fluor-3 and 2',7'-dichlorodihydrofluorescein diacetate, respectively. In vivo efficacy of neuroprotection was evaluated using the Mongolian gerbil of ischemic brain injury model. Result: Reduction of cell viability by glutamate (5 mM) was significantly suppressed by treatment with ginsenoside Rb2. Phosphorylation of MAPKs, Bax, and nuclear AIF was gradually increased by treatment with 5 mM of glutamate and decreased by co-treatment with Rb2. The occurrence of apoptotic cells was decreased by treatment with Rb2 ($25.7{\mu}M$). Cellular $Ca^{2+}$ and ROS levels were decreased in the presence of Rb2, and in vivo data indicated that Rb2 treatment (10 mg/kg) significantly diminished the number of degenerated neurons. Conclusion: Our results suggest that Rb2 possesses neuroprotective properties that suppress glutamate-induced neurotoxicity. The molecular mechanism of Rb2 is by suppressing the MAPKs activity and AIF translocation.

• Clinical and Experimental Pediatrics
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• 제51권10호
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• pp.1112-1117
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• 2008

목 적: 해마 절편 배양에서 산소-포도당 박탈(oxygen-glucose deprivation, OGD)에 의한 세포 사망과 신경 세포 사멸을 propidium iodide(PI) 섭취, Fluoro-Jade(FJ) 염색, TUNEL 염색, caspase-3 면역형광염색 방법으로 관찰하고자 하였다. 방 법: 생후 7일된 Sprague-Dawley 흰쥐의 해마를 MacIlwain chopper로 $350{\mu}m$ 두께의 절편으로 절단하였다. 해마 절편을 6-well plate의 insert 내의 반 유공(sem-porous) 막 위에서 membrane-interface technique으로 10일 동안 배양하였다. 배양된 해마 절편에 산소-포도당 박탈을 60분 동안 가한 후 재산소-재관류하에 기초 배양액에서 48시간 배양하였다. 재산소-재관류 동안 PI 섭취 형광 정도를 시간에 따라 형광 현미경으로 관찰하고 세포사망 백분율(percent cell death)을 측정하였다. 산소-포도당 박탈 직전과 24 시간 후에 해마 절편을 $15{\mu}m$ 두께로 냉동 절단 후 FJ 염색, TUNEL 염색, caspase-3 면역형광염색을 시행하여 세포 사망을 관찰하였다. 결과: OGD 후 PI 섭취 는 해마 절편의 CA1과 DG에 한정되어있었다. OGD 후 재산소-재관류 동안 6시간에서 48시간까지 PI 섭취 형광 강도는 시간이 증가함에 따라 증가하였다. 세포 사망 백분율은 CA1과 DG에서 모두 OGD 후 재산소-재관류 시간이 증가함에 따라 의미 있게 증가하였다(P<0.05). OGD 후 24시간에 세포 변성을 의미하는 많은 FJ 염색 양성 신경 세포 들이 CA1과 DG에서 관찰되었다. 고배율 confocal laser 현미경으로 관찰한 CA1에서의 신경 세포들 중 일부는 명확한 핵과 돌기를 가지고 있는 것을 보여 주었으며, 다른 신경 세포들은 핵의 분절화, 돌기의 손실 등을 보여 주었다. TUNEL 염색과 caspase-3 염색은 OGD 후 24시간에 CA1과 DA에서 TUNEL 양성 발현을 증가시키고 caspase-3 발현을 증가시켰다. 결 론: 해마 절편 배양에서 산소-포도당 박탈 에 의한 다수의 세포 사망을 관찰할 수 있었다. 사망한 세포 들은 주로 신경 세포의 caspase-3 활성화에 의해 매개된 사멸을 보였다.