• 약학회지
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• 제40권6호
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• pp.705-712
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• 1996

We have previously shown that determination of glucose uptake using ${\alpha}$-methylglucose(${\alpha}$-MG) is very sensitive and rapid parameter for the assessment of loss of cellular fu nction in renal cell line($LLC-PK_1$). The present study was designed to elucidate the mechanism of inhibition of ${\alpha}$-MG uptake and the intracellular site of toxic action of cisplatin(CIS). $LLC-PK_1$ cells were exposed to various concentrations(5 ${\mu}$M-l00 ${\mu}$M) of CIS for 5 hrs or 24 hrs and ${\alpha}$-MG uptake was determined. Mitochondrial function was evaluated by measuring intracellular ATP content and MTT reduction. The activities of marker enzymes for the basolateral membrane(Na$^+$-K$^+$ ATPase) and brush border membrane (alkaline phosphatase: ALP) were also measured. CIS treatment significantly inhibited the ${\alpha}$-MG uptake in a time- and dose-dependent manner above 25 ${\mu}$M for 5 hrs. Intracellular ATP content and MTT reduction were affected by 24 hr-treatment of 50 ${\mu}$M CIS. The activities of Na$^+$-K$^+$ ATPase and ALP were significantly decreased at 10 ${\mu}$M and 5 ${\mu}$M of CIS for 24 hrs, respectively. The incubation with CIS for 5 hrs had no effects on the intracellular ATP content, MTT reduction and the activities of marker enzymes up to 100 ${\mu}$M. These results partly indicate that inhibition of ${\alpha}$-MG uptake by CIS may not be attributed to the disturbance of mitochondrial function or inhibition of the activity of Na$^+$-K$^+$ ATPase and can be resulted from direct effect of CIS on the Na$^+$/glucose cotransporter in brush border membrane. This study shows that additional mechanistic information, indicating the intracellular site of nephrotoxic action, can be gained by coupling the ${\alpha}$-MG uptake and ATP content or the activity of Na$^+$-K$^+$ ATPase.

• 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.

• 생약학회지
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• 제39권4호
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• pp.290-294
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• 2008

The objective of this study is screening the anti-oxidative effects of several plant MeOH extracts against oxidative stress in Neuroblastoma cell. Oxidative stress has been implicated in the pathogenesis of many neurotoxicity, neurodegenerative disorders and cell death. This oxidative stress is generated by ROS (Reactive Oxygen Species) such as nitric oxide, nitrogen dioxide, peroxyl, superoxide ($O_2^-$), hydroxyl, alkoxyl. So, in the present study, we induced oxidative stress by treatment of sodium nitroprusside (2.5 mM) in human neuroblastoma SH-SY5Y cell which was treated samples before 24hr, and cell viability was measured by MTT reduction assay. Of those tested, the extracts of Paeonia japonica (roots), Eucommia ulmoides (炒)(barks), Paeonia japonica (曝乾)(roots), Phyllostachys bambusoides (stems), Polygala tenuifolia (去心, 炒)(roots), Paeonia japonica (roots), Polygala tenuifolia (roots), Machilus thunbergii (barks), Mallotus japonicus (leaves), Poria cocos (whole), Sophora flavescens (roots), Angelica tenuissima (roots), Angelica gigas (當歸尾)(roots) showed anti-oxidative effects[$EC_{50}$<15.20 ${\mu}g$/ml(Carnosine:Positive control)]in dose dependent manner.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2004년도 Annual Meeting of the Korean Society ofApplied Pharmacology
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• pp.124-128
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• 2004

We have assessed amyloid ${\beta}-peptide$ $(A{\beta})-induced$ neurotoxicity in primary neurons and organotypic hippocampal slice cultures (OHC) in rat. Exposing cultured hippocampal and cerebellar granule neurons to $A{\beta}$ resulted in a decrease of MTT reduction, and in destruction of neuronal integrity. Treatment of these neurons with tunicamycin, an inhibitor of N-glycosylation in the endoplasmic reticulum (ER), also decreased MTT reduction in these neurons. S-allyl-L-cysteine (SAC), an active organosulfur compound in aged garlic extract, protected hippocampal but not cerebellar granule neurons against $A{\beta}$- or tunicamycin-induced toxicity. In the hippocampal neurons, protein expressions of casapse-12 and GRP 78 were significantly increased after $A{\beta}_{25-35}$ or tunicamycin treatment. The increase in the expression of caspase-12 was suppressed by simultaneously adding $1{\mu}M$ SAC in these neurons. In contrast, in the cerebellar granule neurons, the expression of caspase-12 was extremely lower than that in the hippocampal neurons, and an increase in the expression by $A{\beta}_{25-35}$ or tunicamycin was not detected. In OHC, ibotenic acid (IBO), a NMDA receptor agonist, induced concentration-dependent neuronal death. When $A{\beta}$ was combined with IBO, there was more intense cell death than with IBO alone. SAC protected neurons in the CA3 area and the dentate gyrus (DG) from the cell death induced by IBO in combination with $A{\beta}$, although there was no change in the CA1 area. Although protein expression of casapse-12 in the CA3 area and the DG was significantly increased after the simultaneous treatment of AI3 and IBO, no increase in the expression was observed in the CA1 area. These results suggest that SAC could protect against the neuronal cell death induced by the activation of caspase-12 in primary cultures and OHC. It is also suggested that multiple mechanisms may be involved in neuronal death induced by AI3 and AI3 in combination with IBO.

• Journal of Korean Neurosurgical Society
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• 제59권3호
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• pp.259-268
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• 2016

Objective : Accumulation of advanced glycation end-products (AGE) and mitochondrial glycation is importantly implicated in the pathological changes of the brain associated with diabetic complications, Alzheimer disease, and aging. The present study was undertaken to determine whether sildenafil, a type 5 phosphodiesterase type (PDE-5) inhibitor, has beneficial effect on neuronal cells challenged with AGE-induced oxidative stress to preserve their mitochondrial functional integrity. Methods : HT-22 hippocampal neuronal cells were exposed to AGE and changes in the mitochondrial functional parameters were determined. Pretreatment of cells with sildenafil effectively ameliorated these AGE-induced deterioration of mitochondrial functional integrity. Results : AGE-treated cells lost their mitochondrial functional integrity which was estimated by their MTT reduction ability and intracellular ATP concentration. These cells exhibited stimulated generation of reactive oxygen species (ROS), disruption of mitochondrial membrane potential, induction of mitochondrial permeability transition, and release of the cytochrome C, activation of the caspase-3 accompanied by apoptosis. Western blot analyses and qRT-PCR demonstrated that sildenafil increased the expression level of the heme oxygenase-1 (HO-1). CoPP and bilirubin, an inducer of HO-1 and a metabolic product of HO-1, respectively, provided a similar protective effects. On the contrary, the HO-1 inhibitor ZnPP IX blocked the effect of sildenafil. Transfection with HO-1 siRNA significantly reduced the protective effect of sildenafil on the loss of MTT reduction ability and MPT induction in AGE-treated cells. Conclusion : Taken together, our results suggested that sildenafil provides beneficial effect to protect the HT-22 hippocampal neuronal cells against AGE-induced deterioration of mitochondrial integrity, and upregulation of HO-1 is involved in the underlying mechanism.

• 생약학회지
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• 제38권1호
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• pp.84-89
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• 2007

The present study describes the preliminary evaluation of the antioxidant activities and the neuroprotective effect of methanolic extracts from Psoralea corylifolia Linne (PCE). The antioxidant activities and neuroprotective effect of the PCE were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), MTT reduction assay, and LDH release assay. TPC, DPPH RSA, and RP of the extract at concentration of 100 ${\mu}g$ was 125.93 ${\mu}g$, 63.81%, 0.138, respectively, and those were concentration dependent. The treatment of PC12 and N18-RE-105 cells with various PCE concentrations under $H_2O_2$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay and LDH release assay. Therefore, these results suggest that PCE could be a new potential candidate as an antioxidant against neuronal diseases.

• 한국식품조리과학회지
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• 제27권4호
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• pp.67-73
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• 2011

This study was performed to assess the neuroprotective effects of methanolic extracts from sweet persimmon peel (PPE) against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. The neuroprotective effects of PPE in N18-RE-105 cells were measured using the MTT reduction assay, LDH release assay, and phase-contrast microscopy. The results of the MTT reduction assay showed that treating cells with 500 ${\mu}g/ml$ PPE resulted in cell viability of 66.9%. Additionally, the morphological changes and the results of the LDH release assay showed that glutamate-induced damage to nerve cells was strongly inhibited by PPE. GSH content of N18-RE-105 cells was 3.5 ${\mu}M$ compared to that of the control, whereas pretreatment with 500 ${\mu}g/ml$ PPE increased GSH content by 4.7 ${\mu}M$. PPE was fractionated with hexane, and that layer had the highest neuroprotective effects in glutamate-stressed N18-RE-105 cells. In conclusion, our data showed that glutamate potentiated the effects of N18-RE-105 cell death by a mechanism involving oxidative stress. Therefore, PPE may be a potential candidate for prevention and therapy of neurodegenerative diseases.

• 동의생리병리학회지
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• 제19권1호
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• pp.81-86
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Psoraleae fructus. The extract was studied for dipheny-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being 0.427 mg/mL. The extract was dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (67.7 %) was blocked by the extract concentration- dependently. Furthermore, the extract also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Psoraleae fructus has potent antioxidant activity.

• 동의생리병리학회지
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• 제19권6호
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• pp.1541-1545
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Corm fructus. The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being $200{\mu}g/mL$. The extract at concentrations of $10-500{\mu}g/ml$ showed dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (63.0%) was blocked by the extract (10, 50, 100, 250 and $500{\mu}g/ml$) concentration-dependently. Furthermore, the extract (50, 100 and $250{\mu}g/ml$) also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Corm fructus has potent antioxidant activity.

• KSBB Journal
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• 제27권4호
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• pp.227-231
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• 2012

Whole plants of Zostera asiatica were extracted twice with acetone/methylene chloride (A+M) and methanol (MeOH) in turn. The combined crude extracts were evaporated in vacuo and then the residue was partitioned between water and methylene chloride. The aqueous layer was fractionated into $H_2O$ and n-butanol and then the organic layer was also fractionated into 85% aq. MeOH and n-hexane, successively. The crude extracts and their solvent fractions were evaluated for their inhibitory effect on growth of human cancer cells AGS, HT-29, MCF-7, and HT-1080 cells by MTT reduction assay. Among samples tested, 85% aq. MeOH and n-hexane fractions showed strong cytotoxic effect against AGS, HT-29, and MCF-7 cells. On the other hand, for HT-1080 cell, 85% aq. MeOH fraction exhibited the strongest cytotoxic effect.