• 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.

• 대한본초학회지
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• 제23권3호
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• pp.19-25
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• 2008

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Thujae Semen(TSW) in PC12 cells. Methods : We performed 2,2-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay, 2,2-azinobis- (3-ethyl-benzothiazoline-6-sulfonic acid(ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of TSW. We also carried out 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay(MTT), reactve oxygen species(ROS) assay, and nitric oxide(NO) assay to examine neuroprotective effects against 6-hydroxydopamine(6-OHDA) in PC12 cells. Results : TSW showed $IC_{50}$ values of 404.3 and 219.9 ${\mu}g/mL$ in DPPH and in ABTS assays, respectively. TSW showed 9.74 ${\mu}g/mL$ of total polyphenol contents. TSW incresed cell viability in a dose dependent manner and it showed protective effect against 6-OHDA neurotoxicity at the concentration of 25-200 ${\mu}g/mL$. Moreover, it recovered 6-OHDA induced cell death at the same concentrations. The extract showed a dose dependent reduction of ROS and NO generation by 6-OHDA. Conclusions : We concluded that TSW has neuroprotective effect against 6-OHDA-induced toxicity in PC12 cells through ROS and NO inhibition.

• Nutrition Research and Practice
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• 제7권4호
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• pp.249-255
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• 2013

In this study, the protective effects of EGCG on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced oxidative cell death in catecholaminergic PC12 cells, the in vitro model of Parkinson's disease, were investigated. Treatment with L-DOPA at concentrations higher than $150{\mu}M$ caused cytotoxicity in PC12 cells, as determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry detection. The apoptotic ratio was similar in cells treated with $100{\mu}M$ EGCG plus $150{\mu}M$ L-DOPA (5.02%) and the control (0.96%) (P > 0.05), and was lower than that of cells treated with L-DOPA only (32.24%, P < 0.05). The generation level of ROS (% of control) in cells treated with EGCG plus L-DOPA was lower than that in cells treated with L-DOPA only (123.90% vs 272.32%, P < 0.05). The optical density in production of TBARS in cells treated with L-DOPA only was higher than that in the control ($0.27{\pm}0.05$ vs $0.08{\pm}0.04$, P < 0.05), and in cells treated with EGCG only ($0.14{\pm}0.02$, P < 0.05), and EGCG plus L-DOPA ($0.13{\pm}0.02$, P < 0.05). The intracellular level of GSH in cells treated with EGCG plus L-DOPA was higher than that in cells treated with L-DOPA only ($233.25{\pm}16.44$ vs $119.23{\pm}10.25$, P < 0.05). These results suggest that EGCG protects against L-DOPA-induced oxidative apoptosis in PC12 cells, and might be a potent neuroprotective agent.

• Journal of Microbiology and Biotechnology
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• 제31권4호
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• pp.584-591
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• 2021

Marine algae (seaweed) encompass numerous groups of multicellular organisms with various shapes, sizes, and colors, and serve as important sources of natural bioactive substances. The brown alga Ecklonia cava Kjellman, an edible seaweed, contains many bioactives such as phlorotannins and fucoidans. Here, we evaluated the antioxidative, neuroprotective, and anti-apoptotic effects of E. cava extract (ECE), E. cava phlorotannin-rich extract (ECPE), and the phlorotannin dieckol on neuronal PC-12 cells. The antioxidant capacities of ECPE and ECE were 1,711.5 and 1,050.4 mg vitamin C equivalents/g in the ABTS assay and 704.0 and 474.6 mg vitamin C equivalents/g in the DPPH assay, respectively. The dieckol content of ECPE (58.99 mg/g) was approximately 60% higher than that of ECE (36.97 mg/g). Treatment of PC-12 cells with ECPE and ECE increased cell viability in a dose-dependent manner. Intracellular oxidative stress in PC-12 cells due to ECPE and ECE decreased dose-independently by up to 63% and 47%, respectively, compared with the stress control (323%). ECPE reduced the production of the pro-apoptotic proteins Bax and caspase-3 more effectively than ECE. Early and late apoptosis in PC-12 cells were more effectively decreased by ECPE than ECE treatments. From the results obtained in this study, we concluded that ECPE, which is rich in phlorotannins, including the marker compound dieckol, may be applied to the development of functional materials for improving cognition and memory.

• International Journal of Oral Biology
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• 제45권4호
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• pp.179-189
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• 2020

Green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) is a potent antioxidant with protective effects against neurotoxicity. However, it is currently unclear whether EGCG protects neuronal cells against radiation-induced damage. Therefore, the objective of this study was to investigate the effects of EGCG on ultraviolet (UV)-induced oxidative stress and apoptosis in PC12 cells. The effects of UV irradiation included apoptotic cell death, which was associated with DNA fragmentation, reactive oxygen species (ROS) production, enhanced caspase-3 and caspase-9 activity, and poly (ADP-ribose) polymerase cleavage. UV irradiation also increased the Bax/Bcl-2 ratio and mitochondrial pathway-associated cytochrome c expression. However, pretreatment with EGCG before UV exposure markedly decreased UV-induced DNA fragmentation and ROS production. Furthermore, the UV irradiation-induced increase in Bax/Bcl-2 ratio, cytochrome c upregulation, and caspase-3 and caspase-9 activation were each ameliorated by EGCG pretreatment. Additionally, EGCG suppressed UV-induced phosphorylation of p38 and rescued UV-downregulated phosphorylation of ERK. Taken together, these results suggest that EGCG prevents UV irradiation-induced apoptosis in PC12 cells by scavenging ROS and inhibiting the mitochondrial pathways known to play a crucial role in apoptosis. In addition, EGCG inhibits UV-induced apoptosis via JNK inactivation and ERK activation in PC12 cells. Thus, EGCG represents a potential neuroprotective agent that could be applied to prevent neuronal cell death induced by UV irradiation.

• Bulletin of the Korean Chemical Society
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• 제29권4호
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• pp.833-841
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• 2008

Principal component-genetic algorithm-multiparameter linear regression (PC-GA-MLR) and principal component-genetic algorithm-artificial neural network (PC-GA-ANN) models were applied for prediction of melting point for 323 drug-like compounds. A large number of theoretical descriptors were calculated for each compound. The first 234 principal components (PC’s) were found to explain more than 99.9% of variances in the original data matrix. From the pool of these PC’s, the genetic algorithm was employed for selection of the best set of extracted PC’s for PC-MLR and PC-ANN models. The models were generated using fifteen PC’s as variables. For evaluation of the predictive power of the models, melting points of 64 compounds in the prediction set were calculated. Root-mean square errors (RMSE) for PC-GA-MLR and PC-GA-ANN models are 48.18 and $12.77{^{\circ}C}$, respectively. Comparison of the results obtained by the models reveals superiority of the PC-GA-ANN relative to the PC-GA-MLR and the recently proposed models (RMSE = $40.7{^{\circ}C}$). The improvements are due to the fact that the melting point of the compounds demonstrates non-linear correlations with the principal components.

• 동의생리병리학회지
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• 제17권2호
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

• 생명과학회지
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• 제29권2호
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• pp.173-180
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• 2019

Amyloid ${\beta}$-단백질($A{\beta}$)은 알츠하이머 질병의 특징인 노인성 반점의 주요 성분이며 in vivo와 in vitro에서 신경세포를 대상으로 독성효과를 유발한다. 항산화물질과 프로테오글리칸을 포함한 많은 환경인자들에는 $A{\beta}$의 독성을 완화하는 물질들이 존재한다. 특히, 천연물질들 중에서 자신은 독성이 없으며, 알츠하이머 환자에게 치료효능을 나타내는 천연화합물들을 순수 분리하는 것은 매우 가치가 있다. 본 연구에서는 소엽맥문동의 메탄올 추출물로부터 에틸아세테이트 유기용매로 분획한 물질(OJEA)을 대상으로 in vitro상에서 신경세포독성 제어효능을 탐색하였다. 본 실험을 위해 PC12 세포주에 $A{\beta}_{25-35}$로 유발한 독성에 대한 OJEA 분획물의 억제효능을 MTT 환원법 분석으로 측정하였으며, ${\beta}$-secretase 활성에 대한 OJEA 분획물의 억제효능을 세포기반 ${\beta}$-secretase assay system으로 측정하였다. 또한 PC12 세포에서 $A{\beta}_{25-35}$에 의해 유도된 산화적 스트레스에 대한 OJEA 분획물의 억제효과를 지질과산화 분석법으로 수행하였다. 본 연구의 결과는 OJEA 분획물이 PC12 세포에서 $A{\beta}_{25-35}$에 의해 유도된 세포독성을 강하게 예방 또는 억제하는 효과가 있음을 확인하였으며, 또한 ${\beta}$-secretase의 활성을 억제함으로써 $A{\beta}$의 생성을 완화하는 효과를 예상할 수 있었다. OJEA 분획물은 또한 PC12 세포에서 $A{\beta}_{25-35}$에의 노출에 의하여 유도되는 malondialdehyde (MDA)의 생성을 강하게 억제하였다. 결론적으로, 본 연구의 결과에 의하면 OJEA 분획물에는 $A{\beta}$ 독성에 대한 신경세포의 보호효능을 함유하는 생리활성물질이 함유되어 있음을 제시한다.

• 한국지반공학회논문집
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• 제27권12호
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• pp.69-84
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• 2011

본 연구에서는 NATM Composite 라이닝 공법을 실제 터널 현장에 도입하기 위한 적용성 연구를 수행하였다. 먼저 고품질 PC 패널을 생산하기 위한 최적의 증기양생 조건을 도출하였으며 터널 화재 시 피해를 최소화하기 위해 화재모형 실험을 수행하여 PC 패널 라이닝의 화재 거동 특성을 고찰하였다. 또한 실제 현장에 시험시공을 수행하여 NATM Composite 라이닝 공법의 시공성과 안정성을 확인하였고, 현장 배수실험을 실시하여 경량기포 모르타르의 배수성능을 검토하였다. 그리고 추후 NATM Composite 라이닝 터널 설계 시 효율적으로 PC 패널 라이닝 수량을 산출 할 수 있도록 PC 패널 최적 조합 프로그램을 개발하였으며 공사비 분석 및 VE 평가를 통해 NATM Composite 라이닝 공법의 경제성을 검토하였다. 본 연구 결과 NATM Composite 라이닝 공법은 국내 터널 현장에 충분한 적용성을 갖는 것으로 판단된다.

• The Korean Journal of Physiology and Pharmacology
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• 제12권5호
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• pp.275-280
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• 2008

A brief ischemic insult induces significant protection against subsequent massive ischemic events. The molecular mechanisms known as preconditioning (PC)-induced ischemic tolerance are not completely understood. We investigated whether kinetic changes of cyclooxygenase (COX)-2 during reperfusion time-periods after PC were related to ischemic tolerance. Rats were given PC by occlusion of middle cerebral artery (MCAO) for 10 min and sacrificed after the indicated time-periods of reperfusion (1, 2, 4, 8, 12, 18 or 24 h). In PC-treated rats, focal ischemia was induced by occlusion of MCA for 24 h and brain infarct volume was then studied to determine whether different reperfusion time influenced the damage. We report that the most significant protection against focal ischemia was obtained in rats with 8 h reperfusion after PC. Administration of indomethacin (10 mg/kg, oral) or rofecoxib (5 mg/kg, oral) 48 h prior to PC counteracted the effect of PC. Immunohistochemical analysis showed that COX-2 and HO-l protein were induced in PC-treated rat brain, which was significantly inhibited by rofecoxib. Taken together, we concluded that the kinetic changes of COX-2 expression during the reperfusion period after PC might be partly responsible for ischemic tolerance.