• 한국식품과학회지
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• 제3권1호
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• pp.37-43
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• 1971

(1) 감미종(甘味種) 소형(小型)고추인 Zairaisisi 종자(種子)의 polyphenol oxidase는 최적(最適) pH가 6.5이고 $0{\circ}C$에서도 상당(相當)한 활성(活性)을 가지며 o-diphenol에 대(對)하여 강(强)한 특이성(特異性)을 가지나 monophenol에 대(對)해서는 특이성(特異性)이 없었다. (2) 저온장해(低溫障害)에 의한 갈변초기(褐變初期)에 polyphenol oxidase는 강(强)한 활성도(活性度)를 나타내지만 전면갈변(全面褐變)에 따라 활성도(活性度)는 0 이 되었다. (3) polyphenol 추출액(抽出液)에 상당(相當)하는 효소액(酵素液)의 갈변능(褐變能)을 측정(測定)하였든 바 polyphenol oxidase의 활성도(活性度)가 0 일 때도 in vitro에서는 갈변현상(褐變現象)이 진행(進行)되었고 이때 peroxidase는 당초(當初)와 거의 같은 활성도(活性度)를 지니고 있었다. (4) 저온장해(低溫障害)에 따른 저온초기(低溫初期)에서는 DNP 첨가실험(添加實驗)으로 미루어 호흡(呼吸)의 전자전달계(電子傳達系)에 이상(異常)을 볼 수 있었고 cytochrome oxidase 활성도(活性度)가 저하(低下)하였지만 호흡량(呼吸量)에는 큰 변동(變動)이 생기지 않았기에 조직(組織)호흡(呼吸)에 미치는 chlorogenic acid 첨가실험(添加實驗)을 하여 polyphenol oxidase가 대상호흡계(代償呼吸系)의 terminal oxidase로 쓰인다는 것을 추정(推定)할 수 있었다.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.17-22
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• 2011

Purpose: Nitric oxide (NO) has been known as an important regulator of osteoblasts and periodontal ligament cell activity. This study was performed to investigate the relationship between NO-mediated cell death of human periodontal ligament fibroblasts (PDLFs) and N-methyl-D-aspartic acid (NMDA) receptor antagonist (+)-5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine hydrogen maleate (MK801). Methods: Human PDLFs were treated with various concentrations (0 to 4 mM) of sodium nitroprusside (SNP) with or without $200\;{\mu}M$ MK801 in culture media for 16 hours and the cell medium was then removed and replaced by fresh medium containing MTS reagent for cell proliferation assay. Western blot analysis was performed to investigate the effects of SNP on the expression of Bax, cytochrome c, and caspase-3 proteins. The differences for each value among the sample groups were compared using analysis of variance with 95% confidence intervals. Results: In the case of SNP treatment, as a NO donor, cell viability was significantly decreased in a concentration-dependent manner. In addition, a synergistic effect was shown when both SNP and NMDA receptor antagonist was added to the medium. SNP treated PDLFs exhibited a round shape in culture conditions and were dramatically reduced in cell number. SNP treatment also increased levels of apoptotic marker protein, such as Bax and cytochrome c, and reduced caspase-3 in PDLFs. Mitogen-activated protein kinase signaling was activated by treatment of SNP and NMDA receptor antagonist. Conclusions: These results suggest that excessive production of NO may induce apoptosis and that NMDA receptor may modulate NO-induced apoptosis in PDLFs.

• 한국발생생물학회지:발생과생식
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• 제15권2호
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• pp.113-120
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• 2011

BCL-2 family 단백질들은 세포사멸 신호전달 체계에서 중추적인 역할을 수행하는 것으로 알려져 있으며, BCL2L10 단백질은 그 중 하나로 세포의 사멸과 생존을 조절하는 것으로 알려져 있다. 특이하게도 BCL2L10 단백질은 세포 또는 조직 특이적으로 서로 상반되는 친 세포사멸 또는 항 세포사멸 효과가 각각 보고되어 있다. 현재까지 난소세포에서의 BCL2L10의 발현 여부 및 기능은 알려져 있지 않다. 따라서 본 연구에서 인간 난소 과립세포주인 KGN 세포에서의 BCL2L10 단백질의 발현 여부를 확인한 결과, 상당한 양의 단백질이 발현함을 확인할 수 있었으며, 또한 세포사멸효과를 확인하기 위해서 BCL2L10 단백질을 dose-dependent하게 과발현시킨 후 세포의 생존에 미치는 영향을 분석한 결과,BCL2L10은 KGN 세포에서 과발현 시 세포사멸을 유도함을 관찰하였다. BCL2L10 단백질을 과발현 시 Caspase 9와 3를 활성화 하였으며, 면역염색법을 통해서 BCL2L10 단백질이 미토콘드리아에 위치하는 것을 확인하였다. 또한 BCL2L10단백질의 과발현에 의해 미토콘드리아에서 cytochrome c가 세포질로 분비되는 것을 확인하였다. 이상의 결과로서 본 연구는 BCL2L10의 과발현이 KGN 세포에서 세포사멸을 유도하고, 또한 미토콘드리아에 위치하여 세포질로 cytochrome c를 분비하여 Caspase 9와 3을 활성화 시키는 메커니즘으로 세포사멸을 유도함을 확인하였다. 이러한 연구결과는 BCL2L10단백질이 인간 난소과립세포의 생존과 사멸을 조절하는 인자임을 최초로 규명한 것으로서, 추후 난소에서의 BCL2L10단백질의 생리적인 기능 및 신호 조절 연구의 기반 데이터로서 그 의의가 있으며 활용될 수 있다.

• 대한본초학회지
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• 제21권2호
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• pp.37-46
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• 2006

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8503-8512
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• 2016

The purpose of this study was to provide a detailed explanation of the mechanism of bisanthracycline, WP 631 in comparison to doxorubicin (DOX), a first generation anthracycline, currently the most widely used pharmaceutical in clinical oncology. Experiments were performed in SKOV-3 ovarian cancer cells which are otherwise resistant to standard drugs such as cis-platinum and adriamycin. As attention was focused on the ability of WP 631 to induce apoptosis, this was examined using a double staining method with Annexin V and propidium iodide probes, with measurement of the level of intracellular calcium ions and cytosolic cytochrome c. The western blotting technique was performed to confirm PARP cleavage. We also investigated the involvement of caspase activation and DNA degradation (comet assay and immunocytochemical detection of phosphorylated H2AX histones) in the development of apoptotic events. WP 631 demonstrated significantly higher effectiveness as a pro-apoptotic drug than DOX. This was evident in the higher levels of markers of apoptosis, such as the externalization of phosphatidylserine and the elevated level of cytochrome c. An extension of incubation time led to an increase in intracellular calcium levels after treatment with DOX. Lower changes in the calcium content were associated with the influence of WP 631. DOX led to the activation of all tested caspases, 8, 9 and 3, whereas WP 631 only induced an increase in caspase 8 activity after 24h of treatment and consequently led to the cleavage of PARP. The lack of active caspase 3 had no outcome on the single and double-stranded DNA breaks. The obtained results show that WP 631 was considerably more genotoxic towards the investigated cell line than DOX. This effect was especially visible after longer times of incubation. The above detailed studies indicate that WP 631 generates early apoptosis and cell death independent of caspase-3, detected at relatively late time points. The observed differences in the mechanisms of the action of WP631 and DOX suggest that this bisanthracycline can be an effective alternative in ovarian cancer treatment.

• 대한본초학회지
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• 제21권4호
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• pp.27-36
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• 2006

Objectives : This study was performed for the investigation of anticancer effects of methanol extract of Rheum Rhizoma (MeOH-RR) on a human liver cancer cell line (HepG2). Methods : To study the cytotoxic effect of MeOH-RR on HepG2 cells, the cell viability was determined by XTT reduction method and trypan blue exclusion assay. The cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, MeOH-RR-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome c from mitochondria to cytosol, the level of Bcl-2 and Bax were examined by western blot analysis. Results : MeOH-RR reduced proliferation of HepG2 cells in a dose-dependent manner at 24 h and 48 h treatment. MeOH-RR induced the activation of caspase-3, -8, and -9 and the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3. Furthermore, treatment with MeOH-RR resulted in internucleosomal DNA fragmentation, evidenced by the formation of a DNA ladder on agarose gel, a hallmark of cells undergoing apoptosis. MeOH-RR downregulated Bcl-2, upregulated Bax, and increased the release of cytochrome c from the mitochondria into cytosol in a dose-dependent manner. Moreover, MeOH-RP increased caspase-3 activity. Conclusion : There results suggest that MeOH-RR induce apoptosis via mitochondrial pathway and caspase-3-dependent pathway in HepG2 cells. There results suggest that MeOH-RR is potentially useful as a chemotherapeutic agent in human liver cancer.

• Toxicological Research
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• 제20권3호
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• pp.195-203
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• 2004

4-Tert-Octylphenol (OP) is a surfactant additive widely used in the manufacture of a variety of detergents and plastic products. OP can disrupt endocrine function in humans and animals. This study was carried out to obtain toxicokinetic parameters of OP in male Sprague-Dawley (SD) rats. Male rats were administered with OP by single oral application of 200 mg/kg body weight. Blood, urine and tissues samples were taken at several time intervals after administration. Analysis of samples for OP was performed by column-switching high performance liquid chromatography (HPLC). In addition, we exam-ined tissue distribution and accumulation of OP after single oral application of 50, 100, and 200 mg/kg, single intravenous injection of 1, 5 and 10 mg/kg or daily application of 50 mg/kg for 14 consecutive days. After single oral administration of 200 mg/kg, Cmax of 213 $\pm$ 123 ng/ml was reached within the first 1.3 hr (Tmax) in the plasma. AUC was calculated for 1,333$\pm$484 ngㆍhr/ml. The final elimination half-life of plasma was longer than that of urine, but urinary clearance was lower than oral. A very small fraction of OP (Fe < 0.0017%) was excreted in urine within 24 hr. These results indicated that the major excretion route of OP was not urine. The mean maximal tissue distribution of OP was obserbed at 6 hr after treatment and slowly decreased time-dependently. High OP concentrations were detected in fat at 24 hr. The OP in fat was slowly released with longer elimination half-life and lower clearance than that of other tissues. OP was not accumulated in the liver following single oral application but 14-day oral treatments resulted in two-fold accumulation. It was probably due to the saturation of detoxification pathways. On the other hand, the mRNA expression of cytochrome P450 isoforms except CYP2C11 was not affected by OP at any dose. The expression of CYP2C11 mRNA decreased in a dose-dependent manner. This result suggests that OP changes expression of the male-specific cytochrome P450 isoforms in rat liver, and these changes are closely related to the toxic and estrogenic effect of OP.

• 생명과학회지
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• 제25권11호
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• pp.1235-1243
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• 2015

황흑산은 동의보감의 복강과 장옹의 처방을 위해 기록된 처방전으로 오랫동안 사용되어 왔으나, 항암 효능에 대한 구체적인 연구는 전혀 이루어진 바 없다. 본 연구에서는 AGS 인체 위암세포를 대상으로 황흑산 처리에 의한 증식억제와 연관된 apoptosis 유발 및 관련 기전 연구를 수행하였다. AGS 위암세포에 황흑산 추출물을 처리함에 처리 농도 의존적으로 증식이 억제되었으며, 이는 apoptosis 유발과 연관성이 있음을 핵의 형태적 변형과 sub-G1기 세포의 축적 등으로 확인하였다. 황흑산 추출물에 의한 apoptosis 유도에는 pro-apoptotic Bax 단백질의 발현 증가와 anti-apoptotic Bcl-2의 발현 감소 및 미토콘드리아에서 세포질로의 cytochrome c 유리와 연관성이 있었으며, 세포 내 활성산소종(reactive oxygen species, ROS)의 축적을 증가시켰다. 또한 황흑산 추출물에 의한 apoptosis 유발은 caspases (caspase-3, -8 및 -9)의 활성을 증가시켰으며, poly (ADP-ribose)-polymerase 단백질의 단편화를 초래하였다. 그러나 ROS scavenger 및 pan-caspases inhibitor는 황흑산 추출물에 의한 apoptosis의 유발을 거의 완벽하게 억제하였으며, 암세포의 증식억제도 차단하였다. 이상의 결과는 황흑산 추출물에 의한 apoptosis가 ROS 생성 및 caspase 활성 의존적으로 일어남을 의미하는 것으로 황흑산의 항암기전 해석을 이해하고 향후 지속적인 연구를 위한 유용한 자료로 사용될 것이다.

• 한국산림과학회지
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• 제101권3호
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• pp.517-525
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• 2012

본 연구에서는 4년근 산양삼을 4년 숙성된 감식초에 혼입 숙성시킨 후, 5배 희석하여 쥐에게 경구투여하였다. 이후 당질대사 및 지질대사에 영향을 미치는 단백질 발현율을 분석하여 융합소재의 에너지대사율 증대에 의한 비만억제효과에 대하여 검토하였다. 집단은 대조군(CON), 감식초군(PV), 산양삼+감식초 융합소재군(MPV)으로 각각 구분하였다. 체중의 증가율은 PV, MPV가 낮게 나타났다. 글루코스 농도는 PV와 MPV가 낮았으며, 반대로 GLUT-2는 유의하게 높았다. 유리지방산 농도와 CPT-1은 PV, MPV가 높았으며, MPV가 PV에 비하여 높았다. Cytochrome C oxidase는 MPV가 CON에 비하여 높게 나타났다. AMPK, $PPAR-{\gamma}$ 그리고 $PGC1-{\alpha}$는 모두 PV, MPV가 높았으며, MPV가 PV에 비하여 높았다. 이상과 같은 결과는 융합소재의 체열생산반응을 증명하며, 이를 통한 에너지 대사 상승은 비만억제에 효과적으로 사용될 수 있으리라 판단되며, 다양한 후속연구를 통한 검증이 필요할 것이라 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.769-773
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• 2014

Fangchinoline (Fan) inhibits cell proliferation and induces apoptosis in several cancer cell lines. The effects of Fan on cell growth and proliferation in breast cancer cells remain to be elucidated. Here, we show that Fan inhibited cell proliferation in the MDA-MB-231 breast cancer cell line through suppression of the AKT/Gsk-3beta/cyclin D1 signaling pathway. Furthermore, Fan induced apoptosis by increasing the expression of Bax (relative to Bcl-2), active caspase 3 and cytochrome-c. Fan significantly inhibited cell proliferation of MDA-MB-231 cells in a concentration and time dependent manner as determined by MTT assay. Flow cytometry analysis demonstrated that Fan treatment of MDA-MB-231 cells resulted in cell cycle arrest at the G1 phase, which correlated with apparent downregulation of both mRNA and protein levels of both PCNA and cyclin D1. Further analysis demonstrated that Fan decreased the phosphorylation of AKT and GSK-3beta. In addition, Fan up-regulated active caspase3, cytochrome-c protein levels and the ratio of Bax/Bcl-2, accompanied by apoptosis. Taken together, these results suggest that Fan is a potential natural product for the treatment of breast cancer.