• 한국식품영양과학회지
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• 제42권9호
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• pp.1363-1369
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• 2013

본 연구에서는 플라보노이드 계열 중의 하나인 luteolin을 이용하여 TRAIL에 저항성을 가지는 T24 방광암세포에서 TRAIL 저항성 극복 가능성을 조사하였다. 본 연구의 결과에 의하면 luteolin 및 TRAIL 각각 단독 처리 시 세포증식에 전혀 영향을 미치지 못한 농도의 복합 처리 시 세포증식억제 효과가 크게 증가하였음 알 수 있었다. 이러한 증식억제와 연관된 aspoptosis 유도는 caspase-8의 활성화에 의한 tBid의 발현 증가와 pro-apoptotic 인자인 Bax의 발현 증가로 인한 caspase-9 및 -3의 활성화로 이어지는 type II apoptosis에 의한 것이라 추측되며, 이러한 가정은 각각의 caspase 선택적 저해제를 이용하여 재확인 하였다. 본 연구결과는 TRAIL에 저항성을 보이는 암세포에 luteolin이 감수성을 높이는데 효과적일 수 있으며, 암세포에 대한 combination therapy를 위한 기초자료로 활용성이 높을 것으로 사료된다.

• 한국식품영양과학회지
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• 제40권5호
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• pp.649-659
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• 2011

연구에서는 콩다닥냉이 추출물의 항암활성을 조사하기 위하여 잎 및 뿌리의 열수 추출물(WELVL 및 WELVR)이 HCT116 대장암세포의 증식 억제와 연관된 apoptosis 유도 기전에 관한 연구를 시도하였다. 본 연구의 결과에 의하면 HCT116 세포에 WELVL 및 WELVR을 처리하였을 경우에 유발되는 증식 억제 및 형태 변화는 apoptosis 유발과 밀접한 연관이 있었으며, 증식억제 및 apoptosis 유도 효과는 WELVL에 비하여 WELVR에서 높게 나타났다. 특히 WELVR에 의한 apoptosis 유발에는 FasL의 발현 증가를 통한 caspase-8의 활성화와 이로 인한 Bid 단백질의 단편화와 함께 Bcl-2 family의 발현 변화를 통한 mitochondria의 기능 이상과 이로 인한 caspase-9 및 -3의 활성화, 그리고 기질단백질들의 분해가 중요한 역할을 하는 것으로 나타났다. 또한 IAP family의 발현 감소로 인한 caspase의 활성 증가도 어느 정도 관여하는 것으로 생각되어진다. 따라서 WELVR 처리에 의하여 유발되는 apoptosis는 extrinsic pathway 및 intrinsic pathway를 모두 경유하는 multiple apoptotic pathway에 의하여 조절되는 것으로 생각되며, 이러한 결과들은 인체 암세포에서 콩다닥냉이의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 콩다닥냉이 추출물을 포함한 그와 유사한 항암제 후보물질들의 연구 기초자료로서 사용될 수 있을 것으로 생각된다.

• Journal of Korean Neurosurgical Society
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• 제50권5호
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• pp.403-408
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• 2011

Objective : Contrary to some clinical belief, there were quite a few studies regarding animal models of intracerebral hemorrhage (ICH) $in$ $vivo$ suggesting that prior use of statins may improve outcome after ICH. This study reports the effect of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG CoA) reductase inhibitor, simvastatin given before experimental ICH. Methods : Fifty-one rats were subjected to collagenase-induced ICH, subdivided in 3 groups according to simvastatin treatment modality, and behavioral tests were done. Hematoma volume, brain water content and hemispheric atrophy were analyzed. Immunohistochemical staining for microglia (OX-42) and endothelial nitric oxide synthase (eNOS) was performed and caspase-3 activity was also measured. Results : Pre-simvastatin therapy decreased inflammatory reaction and perihematomal cell death, but resulted in no significant reduction of brain edema and no eNOS expression in the perihematomal region. Finally, prior use of simvastatin showed less significant improvement of neurological outcome after experimental ICH when compared to post-simvastatin therapy. Conclusion : The present study suggests that statins therapy after ICH improves neurological outcome, but prior use of statins before ICH might provide only histological improvement, providing no significant impact on neurological outcome against ICH.

• 약학회지
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• 제49권4호
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• pp.355-364
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• 2005

A neurotoxin, 6-hydroxydopamine (6-OHDA) has long been used to form a Parkinson's disease (PD) model by inducing the lesion in catecholaminergic pathways, particularly the nigrostriatal dopamine (DA) pathway. Whereas l-deprenyl, a selective inhibitor of monoamine oxidase (MAO) type B, is now widely used in the treatment of PD, the precise action mechanism of the drug remains elusive. In this study, we investigated whether l-deprenyl shows protective effect against the DA depletion induced by 6-OHDA in rat brain, and against 6-OHDA-induced neurotoxicity and oxidative stress in catecholaminergic neuroblastoma SH-SY5Y cells that are known to lack MAO-B activity. Pretreatment of l-deprenyl significantly enhanced the striatal DA, 3,4-dihydroxyphenylacetic acid, homovanilic acid, and 3-methoxytyramine levels compared to the untreated 6-OHDA-lesioned rat, indicating that l-deprenyl pretreatment prevents 6-OHDA-induced depletion of not only striatal dopamine but also its metabolites. Treatment of 6-OHDA for 24hrs decreased the cell viability and increase the generation of ROS in dose-dependent manners. We further investigated whether caspase activity is involved in the action of l-deprenyl. Treatment of l-deprenyl $(0.1\~100{\mu}M)$ did not produce any changes in 6-OHDA-induced cleavage of poly (ADP-ridose) polymerase in SH-SY5Y cells. Our results suggest that the neuroprotective effect of l-deprenyl against 6-OHDA is due to its increased scavenger activity, but independent of inhibition of MAO-B or caspase-3 activation.

• 동의신경정신과학회지
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• 제20권2호
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• pp.1-17
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• 2009

Objectives : Alzheimer's disease(AD) is the most common form of dementia, which is characterized by progressive deterioration of memory and higher cortical functions that ultimately results in total degradation of intellectual and mental activities. Nokyongdaebo-tang(Lurongdabutang) has been usually used for the treatment for the deficiency syndrome dementia and amnesia. This experiment was designed to investigate the effect of the Nokyongdaebo-tang(Lurongdabutang) hot water extract on pathological AD model. Methods : The effects of the Nokyongdaebo-tang(Lurongdabutang) hot water extract on cultured spinal cord cells induced by ${\beta}$-amyloid were investigated. The effects of the Nokyongdaebo-tan(Lurongdabutang) hot water extract on the memory deficit mice induced by scopolamine were investigated. Results : 1. ${\beta}$-amyloid treatment on cultured spinal cord cells increased both GFAP-staining intensity of astrocytes and caspase 3 immunoreactivity on cultured cells. Then, Nokyongdaebo-tang(Lurongdabutang) treatment reduced the labeling intensity for both GFAP and caspase 3 proteins in culture cells. 2. Scopolamine treatment into mice increased levels of GFAP-positive astrocytes and caspase 3-labeled cells of the hippocampal subfields dentate hilar region, CA3 and CA1 area. In vivo administration of Nokyongdaebo-tang(Lurongdabutang) attenuated labeling intensity for those two proteins in the same hippocampal areas. Similar effects were observed by the treatment of galanthamine, an inhibitor of acetylcholinesterase. Conclusions : This experiment shows that the Nokyongdaebo-tang(Lurongdabutang) may play a protective role in damaged neural tissues. Since neuronal damage seen in degenerative brains such as AD are largely unknown, the current data may provide possible insight into therapeutic strategies for AD treatments. Nokyongdaebo-tang(Lurongdabutang) might be effective for the prevention and treatment of AD.

• 한국환경성돌연변이발암원학회지
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• 제24권3호
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• pp.113-120
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• 2004

Chromium compounds are known human and animal carcinogens. In this study, the effects of sodium chromate on apoptosis and cell cycle were investigated in order to unveil the elements of early cellular responses to the metal. Using Chinese hamster ovary cells(CHO-K1-BH4), we found taht chromium (VI) treatment induced apoptosis in these cells, as signified by nuclear fragmentation, DNA laddering on agarose gel electrophoresis, and an increased proportionof cells with hypodiploid DNA. Preceding these changes, chromium (VI) treatment increased caspase 3 pritease activity and also increased expression of p53 protein, while the level of bcl2 protein was not changed. Coincubation with caspase inhibitor, Z-DEVD-FMK, inhibited chromium-induced apoptosis. In the flow cytometric analysis using propidium iodide fluorescence, an increase of cell population in G2/M phase was shown in cells exposed to at least 160 $\mu\textrm{m}$ of sodium chromate for 72h, form 9.8% for 0$\mu\textrm{m}$ chromium (VI) to 26.4% for 320$\mu\textrm{m}$ chromium(VI). Taken together, these findings suggest that chromium(VI)-induced apoptosis is accompanied by G2/M cell cycle arrest, and that p53-mediated pathway may be involved in positive regulation of G2/M arrest and a concurred apoptosis in CHO cells.

• 대한한의학회지
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• 제21권4호
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• pp.64-72
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• 2000

Objectives : This study was designed to investigate the neuroprotective effect of Hwansodan(Huanshao-dan) on the apoptosis induced by withdrawal of neurotrophic support. Methods : PCl2 pheochromocytoma cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MTT assay. We used DNA fragmentation and caspase 3-like protease activation assay. Results : The water extract of Hwansodan(Huanshao-dan) significantly showed protective effects on serum and glucose deprivation-induced apoptotic death. Hwansodan(Huanshao-dan) also prevents DNA fragmentation and caspase 3-like protease activation, representing typical neuronal apoptotic phenomena in PCl2 pheochromocytoma cells and induces tyrosine phosphorylation of proteins around 44 kDa, which was identified as ERK1 with electrophoretic gel mobility shift by Western blot. In addition, MAPK kinase(MEK) inhibitor PD98059 and Ras inactivator, ${\alpha}-hydroxyfarnesylphosphonic$ acid attenuated the neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Conclusions : These results indicate that Ras/MEK/ERK signaling pathway plays a key role in neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Taken together, we suggest the possibility that Hwansodan(Huanshao-dan) might provide a neurotrophic-like activity in PCl2 cells.

• Journal of Life Science
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• 제10권2호
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• pp.39-44
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• 2000

Trichostatin A (TSA) is a Streptomyces product, which inhibits the enzyme activity of histone deacetylase. It is also known as an inducer of apoptosis in several human cancer cell lines. In this study, we investigated the mechanism of apoptosis induced by TSA in MDA-MB-231 human breast carcinoma cells. The cytotoxicity of TSA on MDA-MB-231 cells was assessed by MTT assay. The cell viability was decreased dose-dependently and the IC\ulcorner value was about 100 ng/ml after 48 h treatment with TSA. Morphological change and DNA ladder formation, the biochemical hallmarks of apoptotic cell death, were observed after treatment of TSA in a concentration-dependent manner, which was accompanied with cleavage of poly(ADP-ribose) polymerase and $\beta$-catenin, and activation of caspase-3. TSA treatment up-regulated the expression of a cyclin-dependent kinase inhibitor p21 (Wafl/Cip1) protein, a key regulatory protein of the cell cycle. However, there is no detectable change of both Bcl-2 and Bax expressions. These results demonstrated that TSA might inhibit cell growth through apoptosis in human breast carcinoma MDA-MB-231 cells.

• 한국자원식물학회지
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• 제33권4호
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• pp.279-286
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• 2020

본 연구에서는 부처꽃 에탄올 추출물(ELM)에 대한 항암효능을 알아보기 위하여 인체백혈병 U937 세포의 증식에 미치는 영향과 이와 연관된 apoptosis 유발 여부와 함께 그에 따른 분자생물학적 기전에 대해서 조사하였다. 먼저 ELM 처리에 따른 증식 억제 정도를 조사한 결과, ELM 처리 농도 의존적으로 생존율 및 증식억제 현상이 나타났으며, 핵의 형태 변화, DNA 단편화 및 apoptosis 유발에 관하여 조사한 결과 역시 ELM 처리 농도 의존적으로 증가됨을 확인할 수 있었다. ELM 처리에 따른 U937 세포에서의 apoptosis 유발에 있어서 미토콘드리아 막의 기능 손상이 관여하는 지를 확인하기 위하여 MMP의 변화 정도를 확인한 결과, ELM 처리 농도 증가에 따라 MMP의 소실이 증가하는 것을 관찰할 수 있었다. 이러한 MMP의 소실에 가 관여하는 지를 확인하기 위하여 사멸수용체(DR4, 5, Fas) 및 사멸수용체에 결합하는 리간드(FasL, TRAIL)의 발현 변화를 확인한 결과, DR4 및 DR5의 발현이 증가하는 것으로 관찰되었다. 또한 내인적 경로에 관여하는 Bcl-2 family 유전자들의 발현변화를 확인한 결과, Bcl-2 발현 감소 및 Bax의 발현 증가의 변화를 보였으며, Bid 단백질의 발현감소가 나타났으므로 상대적으로 tBid의 생성이 증가되었음을 추측할 수 있었다. 한편apoptosis 유발에 직접적으로 관여하는 것으로 알려진 caspase-3, -8 및 -9의 발현에 미치는 ELM의 영향에 대해서 조사하였다. 결과에서 알 수 있듯이 ELM은 death receptor에 의하여 활성화 되는 것으로 알려진 caspase-8 및 세포질로 방출된 cytochrome c에 의하여 활성화 되는 것으로 알려진 caspase-9의 활성화를 유발하였으며, caspase cascade에 의하여 apoptosis에 직접적으로 관여하는 caspase-3의 발현도 증가시키는 것으로 나타났다. 또한 활성화된 caspase-3에 의하여 분해가 일어나는 기질 단백질인 PARP의 경우 ELM 처리에 의하여 모두 단편화가 유발되는 것으로 나타났다. 이상의 결과를 종합해 보면 인체 백혈병 U937 세포에 ELM을 처리하였을 경우에 유발되는 apoptosis는 외인적 경로인 DR4 및 DR5의 발현 증가를 통한 caspase-8의 활성화와 이로 인한 Bid 단백질의 단편화와 함께 내인적 경로의 미토콘드리아 기능 손실에 의하여 caspase-9 및 -3의 활성화 유발과 기질단백질들의 분해가 중요한 역할을 하는 것으로 생각되며, IAP family의 발현 감소로 인하여 caspase의 활성이 억제되지 못하는 것도 apoptosis 유도에 어느 정도 관여했을 것으로 생각된다. 따라서 ELM 처리에 의하여 유발되는 apoptosis는 외인적 경로 및 내인적 경로를 모두 경유하는 multiple apoptotic pathway에 의하여 조절되며, 이때 caspases가 중요한 역할을 한다는 것을 알 수 있었다.

• 동의생리병리학회지
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• 제27권1호
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• pp.43-48
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• 2013

Cisplatin is a anti-neoplastic agent which is commonly used for the treatment of solid tumor. Cisplatin activates multiple signal transduction pathways involved in the stress-induced apoptosis in a variety of cell types. Previous study reported that cisplatin induces apoptosis through activation of ERK, p38 and JNK in rat mesangial cells, but apoptotic pathway remain known. The present study investigated the apoptotic pathway for cisplatin-indcued apoptosis in rat mesangial cells. cisplatin-induced apoptosis was associated with activation of caspase-3, caspase-8, caspase-9. Caspase-8 inhibition prevented the activation of both caspase-3 and caspase-9. In addition, cisplatin-induced apoptosis increased the expression of Bax, but not the level of Bcl-2. These change of Bax/bcl-2 ratio caused the release of cytochrome c from mitochondria into cytosol. In previous study, the ethanol extract of Bojungbangam-tang (EBJT) inhibited cisplatin-induced apoptosis in rat mesangial cells through inhibition of ERK and JNK activation. However, EBJT did not increase cell proliferation, because it did not prevent cisplatin-induced G2/M phase arrest. These effect of EBJT may be related to p38 activation. Cisplatin-induced G2/M phase arrest are inhibited by treatment with p38 inhibitor and EBJT in rat mesangial cells. Also, p38 inhibition and EBJT treatment on cisplatin-induced G2/M phase arrest are markedly increased the G0/G1 phase and reduced the sub-G1. In conclusion, anti-apoptotic effet of EBJT did not increases cell proliferation, because EBJT did not reduce p38 activation related to cisplatin-induced G2/M phase arrest.