• 생약학회지
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• 제44권4호
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• pp.379-383
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• 2013

Portulaca oleracea L. is known to have many biological benefits such as anti-oxidant, anti-inflammatory, anti-allergic and anti-tumor. The objective of this study is to explore the neuroprotective effect of P. oleracea L. against glutamate-induced oxidative stress in mouse hippocampal HT22 cells. P. oleracea L. 70% ethanol extract and solvent fractions have the potent neroprotective effects on glutamate-induced nerotoxicity by induced the expression of heme oxygenase (HO)-1 in HT22 cells. Especially, ethyl acetate fraction showed higher protective effect. In HT22 cell, P. oleracea L. treatment with ERK inhibitor (PD98059) and c-JUN N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea L. ethyl acetate fraction induced HO-1 expression and P. oleracea L. ethyl acetate fraction also increased ERK and JNK phosphorylation. Furthermore, we found that treatment of P. oleracea L. caused the nuclear accumulation of Nrf2. In conclusion, the ethyl acetate fraction of 70% ethanol extract of P. oleracea L. significantly protect glutamate-induced oxidative damage by induction of HO-1 via Nrf2, ERK and JNK pathway in mouse hippocampal HT22. Taken together these finding suggest that P. oleracea L. ethyl acetate fraction is good source for taking active compounds and may be a potential therapeutic agent for brain disorder that induced by oxidative stress and neuronal damage.

• International Journal of Oral Biology
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• 제34권1호
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• pp.29-36
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• 2009

Cyclosporin A (CsA) has been used clinically as an immunosuppressive drug to prevent organ transplant rejection and in basic research as a mitochondrial permeability blocker. It has been reported that CsA has a protective role in severed neurons and a neurotrophic effect in neuronal cells. However, the molecular mechanisms underlying the stimulation of neuronal cell proliferation by CsA have not yet been elucidated. In our current study, we investigated CsA responsive proteins in PC12 cells using a systematic proteomic approach. The viability of these cells following CsA treatment increased in a dose- and time-dependent manner. Proteins in the CsA-treated PC12 cells were profiled by two-dimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) and electrospray ionization quadupole time-of-flight mass spectrometries (EIQ-TOFMS). This differential expression analysis showed significant changes for 10 proteins (6 up-regulated and 4 down-regulated) upon CsA treatment that were related to cell proliferation, metabolism and the stress response. These proteomics data further our understanding of the proliferation mechanisms of PC12 cells exposed to CsA and demonstrate that our methodology has potential to further elucidate the mechanisms and pathways involved.

• 한국식품저장유통학회지
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• 제18권1호
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• pp.124-129
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• 2011

PC12 신경세포를 이용하여 구아바 열매와 잎 추출물이 $H_2O_2$로 유도된 신경세포 독성에 대한 보호 효과를 조사하였다. 구아바 열매와 잎 열수 추출물의 총페놀성 화합물 함량은 각각 11.75 및 293.25 mg/g이었고, gallic acid 함량은 각각 22.78 및 117.34 mg/100 g이었다. $H_2O_2$ 처리한 PC12 cell내의 활성산소 생성억제효과를 측정한 결과 구아바 잎열수 추출물에서 높은 활성산소 생성 억제효과를 보였다. MTT방법을 이용하여 $H_2O_2$로 유도된 PC12 신경세포에 대한 보호효과를 측정한 결과 구아바 열매와 잎 열수 추출물은 높은 신경세포 보호효과를 보였고, LDH release 실험결과 모든 농도에서 세포막 보호효과를 나타내었다. 본 연구 결과를 종합해 볼 때, 구아바 열매와 잎 열수 추출물의 항산화활성과 산화적 스트레스로부터의 신경세포 보호효과를 나타내어 퇴행성 뇌신경질환 등의 예방 및 개선을 위한 산업화 소재로서의 활용 가능성이 높다고 판단된다.

• 동의생리병리학회지
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• 제18권4호
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• pp.1071-1077
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• 2004

Mudanpi (Cortex Moutan Radicis; the root cortex of Paeonia suffruticosa Andrews) is an important Chinese crude drug used in many oriental prescriptions. 1,2,3,4,6-Penta-O-galloyl-beta-D-glucose (PGG), a major component of this crude drug, has been shown to possess potent antioxidant, anti-mutagenic and anti-proliferative effects. In this study, I examined whether PGG could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 expression and HO activity. Exposure of Neuro 2A cells to PGG (10-50μM) resulted in a concentration- and time-dependent induction of HO-1 mRNA, and protein expressions and heme oxygenase activity. PGG protected the cells from hydrogen peroxide-induced cell death. The protective effect of PGG on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX (ZnPP IX), a HO inhibitor. These results indicate that PGG is a potent inducer of HO-1 and HO-1 induction is responsible for the PGG-mediated cytoprotection against oxidative damage.

• 대한한방내과학회지
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• 제25권3호
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• pp.461-472
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• 2004

Objectives : For the screening of neuroprotective effects of medicinal herbs, the complex system of animal models suffer some disadvantages in controlling critical parameters such as blood pressure and body temperature. Additionally, application of drugs to the appropriate brain area sometimes is difficult, due to poor permeability though the blood brain barrier, and so potential protective effects might be masked. Methods : Organotypic hippocampal slice culture (OHSC) method has the advantages of being relatively easy to prepare and of maintaining the general structure, including tissue integrity and the connections between cells. Drugs can easily be applied and neuronal damage can easily be quantified by using tissues and culture media. This study demonstrates neuroprotective effects of Puerariae radix (葛根, PR), Salviae miltiorrhizae radix (丹蔘, SR), Rhei rhizoma (大黃, RR), and Bupleuri radix (柴胡, BR). These were screenedand compared to MK-801, antagonist of NMDA receptors, by using OHSC of 1 week-old Sprague-Dawley rats. Oxygen/glucose deprivation (OGD) were conducted in an anaerobic chamber $(85%\;N_2,\;10%\;CO_2\;and\;5%\;H_2)$ in a deoxygenated glucose-free medium for 60 minutes. Water extracts of each herbs were treated to culture media with $5\;{\mu}g/ml$ for 48 hours. Results : Neuronal cell death in the cultures was monitored by densitometric measurements of the cellular uptake of propidium iodide (PI). PI fluorescence images were obtained at 48 hours after the OGD and medicinal herb treatment. Also TUNEL-positive cells in the CAI and DG regions and LDH concentrations in culture media were measured at 48 hours after the OGD. According to measured data, MK-801, PR, SR and BR demonstrated significant neuroprotective effect against excessive neuronal cell death and apoptosis induced by the OGD insult. Especially, PR revealed similar neuroprotective effect to MK-801 and RR demonstrated weak neuroprotective effect. Conclusions : These results suggest that OHSC can be a suitable method for screening of neuroprotective effects of medicinal herbs. (This work was supported by the research program of Dongguk University and Grant 01-PJ9-PG1-01CO03-0003 from Ministry of Health & Welfare.)

• Journal of Magnetics
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• 제19권4호
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• pp.357-364
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• 2014

This study examined effect of a transcranial magnetic stimulation device with a commercial-frequency approach on the neuronal cell death caused ischemia. For a simple transcranial magnetic stimulation device, the experiment was conducted on an ischemia induced rat by transcranial magnetic stimulation of a commercial-frequency approach, controlling the firing angle using a Triac power device. The transcranial magnetic stimulation device was controlled at a voltage of 220 V 60 Hz and the trigger of the Triac gate was varied from $45^{\circ}$ up to $135^{\circ}$. Cerebral ischemia was caused by ligating the common carotid artery of male SD rats and reperfusion was performed again to blood after 5 minutes. Protein Expression was examined by Western blotting and the immune response cells reacting to the antibodies of Poly ADP ribose polymerase in the cerebral nerve cells. As a result, for the immune response cells of Poly ADP ribose polymerase related to necrosis, the transcranial magnetic stimulation device suppressed necrosis and had a protective effect on nerve cells. The effect was greatest within 12 hours after ischemia. Therefore, it is believed that in the case of brain damage caused by ischemia, the function of brain cells can be restored and the impairment can be improved by the application of transcranial magnetic stimulation.

• The Korean Journal of Physiology and Pharmacology
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• 제15권6호
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• pp.353-361
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• 2011

In this study, cyanidin-3-glucoside (C3G) fraction extracted from the mulberry fruit (Morus alba L.) was investigated for its neuroprotective effects against oxygen-glucose deprivation (OGD) and glutamate-induced cell death in rat primary cortical neurons. Cell membrane damage and mitochondrial function were assessed by LDH release and MTT reduction assays, respectively. A time-course study of OGD-induced cell death of primary cortical neurons at 7 days in vitro (DIV) indicated that neuronal death was OGD duration-dependent. It was also demonstrated that OGD for 3.5 h resulted in approximately 50% cell death, as determined by the LDH release assay. Treatments with mulberry C3G fraction prevented membrane damage and preserved the mitochondrial function of the primary cortical neurons exposed to OGD for 3.5 h in a concentration-dependent manner. Glutamate-induced cell death was more pronounced in DIV-9 and DIV-11 cells than that in DIV-7 neurons, and an application of $50{\mu}M$ glutamate was shown to induce approximately 40% cell death in DIV-9 neurons. Interestingly, treatment with mulberry C3G fraction did not provide a protective effect against glutamate-induced cell death in primary cortical neurons. On the other hand, treatment with mulberry C3G fraction maintained the mitochondrial membrane potential (MMP) in primary cortical neurons exposed to OGD as assessed by the intensity of rhodamine-123 fluorescence. These results therefore suggest that the neuroprotective effects of mulberry C3G fraction are mediated by the maintenance of the MMP and mitochondrial function but not by attenuating glutamate-induced excitotoxicity in rat primary cortical neurons.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2006년도 Spring Conference
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• pp.123-135
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• 2006

Alzheimer's disease (AD) is the most common form of dementia in the aging population and is clinically characterized by a progressive loss of cognitive abilities. Pathologically, it is defined by the appearance of senile plaques - extracellular insoluble, congophilic protein aggregates composed of amyloid $\beta$ (A$\beta$) and neurofibrillary tangles (NFTs) - inyracellular lesions consisting of paired helical filaments from hyperphosphorylated cytoskeletal tau protein as described by Alois Alzheimer a century ago. These hallmarks still serve as the major criteria for a definite diagnosis of the disease. Consequently, one of the key strategy for drug development in this disease area focuses on reducing the concentration of cerebral A$\beta$ plaque by using substances that inhibit A$\beta$ fibril formation. We focused on developing inhibitors by synthesizing several kinds of aromatic molecules. The synthetic compounds were initially screened to evaluate the effective compound by tioflavin T fluorescence assay. The selected effective compounds were tested cytotoxicity and protective effect from A$\beta$-induced neuronal toxicity by cell based MTT assay with HT22 hippocampal neurons. The BBB permeability on effectors was also tested in in vitro co-culture model(HUVEC/C6 cell line). The behavior test wea carried out in mutant APP/PS1 transgenic mouse model of Alzheimer's disease. And inhibition of A$\beta$ fibril formation by the effective compound was monitored with transmitted electron microscopic images.

• 대한한의학회지
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• 제23권1호
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• pp.50-60
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• 2002

Objectives: Sunghyangjunggisan (SHJS) is a commonly prescribed drug with a wide neuropharmacological spectrum. The water extracts of SHJS were found to be protective against neurotoxicity elicited by deprivation of serum and glucose. Methods: The morphological examination and Hoechst staining of nucleus also clearly showed that the extracts attenuated the cell shrinkage, membrane blebbing, representing typical neuronal apoptotic phenomena and nucleosome-sized fragmentation under the microscope in PC 12 rat pheochromocytoma cells. Results: Activation of protein kinase A (PKA) with dibutyl-cAMP and forskolin also protected during glucose deprivation, although it was not additive with the effect provided by phorbol ester. Interestingly, treatment with the protein kinase A inhibitor, KT5720, was not neuroprotective in the presence of SHJS. Electrophoretic mobility shift assays were used to characterize the neuroprotective binding of nuclear proteins to consensus sequences for AP-l, nuclear factor kappa B ($NF-{\kappa}B$) after glucose deprivation. When PC 12 cells are induced to undergo apoptosis by serum deprivation, AP-l and $NF-{\kappa}B$ DNA binding activity transiently increases to a slight degree. This stimulation is blocked by the water extracts of SHJS. The site of action of the drugs appeared to involve specific inhibition of AP-1 and nuclear factor kB binding activity. Conclusions: Taken together, these results suggested the possibility that the extracts of SHJS might provide a neurotrophic-like activity in PC 12 cells.

• 생명과학회지
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• 제19권7호
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• pp.963-967
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• 2009

본 연구에서는 glutamate로 유도된 세포독성으로부터 신경세포를 보호하는 상백피(Morus alba) 추출물의 활성을 확인하기 위하여 N18-RE-105 세포주를 이용하여 MTT reduction assay, LDH release assay 및 광학 현미경을 이용하여 형태학적인 변화를 관찰하였다. 그 결과, 상백피 methanol 추출물에서 농도 의존적으로 신경세포 보호효과가 나타났으며, 50 본 연구에서는 glutamate 로 유도된 세포독성으로부터 신경세포를 보호하는 상백피 (Morus alba) 추출물의 활성을 확인하기 위하여 NI8-RE-I05 세포주를 이용하여 MTT reduction assay, LDH release assay 및 광학 현 미 경 을 이 용하여 형태학적인 변화를 관찰하였다. 그 결과y 상백피 methanol 추출물에서 농도 의존적으로 신경세포 보호효과가 나타났으며, 50 ${\mu}g$/ml 농도에서는 80% 이상의 세포생존율을 확인할수 있었다. 이 결과는 N18-RE-105 세포주의 LDH release assay와 형태학적 변화에서도 일치하는 결과를 확인하였다. 가장 높은 활성을 보인 상백피 methanol 추출물을 hexane, diethyl ether, ethyl acetate, water 층으로 분획하여 각 각 1, 5, 10 ${\mu}g$/ml 농도로 처리 시 hexane 층에서 48.0%, 65.6%, 71.5%로 가장 높은 신경세포 보호효과를 확인할 수 있었다. 따라서 상백피 추출물은 glutamate 에 의한 세포독성으로부터 신경세포 손상을 억제하며 신경세포를 보호하는 효과가 있다는 것을 알 수 있었다.