• Nutrition Research and Practice
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• 제11권2호
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• pp.97-104
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• 2017

BACKGROUND/OBJECTIVE: Although Angelica keiskei (AK) has widely been utilized for the purpose of general health improvement among Asian, its functionality and mechanism of action. The aim of this study was to determine the protective effect of ethanol extract of AK (AK-Ex) on acute hepatotoxicity induced by acetaminophen (AAP) in HepG2 human hepatocellular liver carcinoma cells and HepaRG human hepatic progenitor cells. MATERIALS/METHODS: AK-Ex was prepared HepG2 and HepaRG cells were cultured with various concentrations and 30 mM AAP. The protective effects of AK-Ex against AAP-induced hepatotoxicity in HepG2 and HepaRG cells were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, lactate dehydrogenase (LDH) assay, flow cytometry, and Western blotting. RESULTS: AK-Ex, when administered prior to AAP, increased cell growth and decreased leakage of LDH in a dose-dependent manner in HepG2 and HepaRG cells against AAP-induced hepatotoxicity. AK-Ex increased the level of Bcl-2 and decreased the levels of Bax, Bok and Bik decreased the permeability of the mitochondrial membrane in HepG2 cells intoxicated with AAP. AK-Ex decreased the cleavage of poly (ADP-ribose) polymerase (PARP) and the activation of caspase-9, -7, and -3. CONCLUSIONS: These results demonstrate that AK-Ex downregulates apoptosis via intrinsic and extrinsic pathways against AAP-induced hepatotoxicity. We suggest that AK could be a useful preventive agent against AAP-induced apoptosis in hepatocytes.

• 대한한의학방제학회지
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• 제30권2호
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• pp.67-83
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• 2022

Objectives : In this study, the anticancer activity of water extracts of three herbal medicine formulas, Bigihwan (BGH), Daechilgitang (DCGT) and Mokwhyangbinranghwan (MHBRH) listed in Donguibogam, was evaluated in HepG2 cells, a human hepatocellular carcinoma cell line. Methods : We investigated whether the cell viability of HepG2 cells was inhibited by the treatment of water extracts of three prescriptions, and whether their viability inhibitory effect was related to the induction of apoptosis. In addition, the role of autophagy on the induction of apoptosis by the treatment of these extracts was investigated. Results : The anticancer activity of the three water extracts on HepG2 cells was due to induction of apoptosis, not necrosis. Among them, BGH activated the caspase-dependent intrinsic apoptosis pathway associated with mitochondrial dysfunction. However, autophagy was induced more than 2-fold in DCGT-treated HepG2 cells, and the anticancer activity of DCGT was enhanced 1.5-fold in the presence of an autophagy inhibitor, but was attenuated in BGH and MHBRH-treated cells. Conclusion : The results of this study indicate that DCGT-induced autophagy was involved in the inhibition of apoptosis, whereas autophagy by BGH and MHBRH was related to induction of apoptosis.

• 생명과학회지
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• 제16권7호
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• pp.1235-1242
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• 2006

브로콜리를 포함한 십자화과 식물에서 glucoraphanin의 가수분해를 통해 생성되는 isothiocyanate의 일종인 sulforaphane은 역학적 조사를 포함한 다양한 선행 연구에서 강력한 암예방 효과를 가지는 것으로 알려져 있다. 항암효과에 관한 최근 연구 결과에 따르면 sulforaphane은 다양한 인체암세포의 증식을 억제하고 apoptosis를 유발할 수 있는 것으로 알려지고 있으나, 정확한 분자생물학적 기전은 밝혀져 있지 않은 상태이다. 본 연구에서는 sulforaphane의 항암작용 기전을 조사하기 위하여 HepG2 인체간암세포의 증식에 미치는 sulforaphane의 영 향을 조사하였다. Sulforaphane의 처리에 의한 HepG2 세포의 증식억제 및 형태적 변형은 세포주기 G2/M arrest 및 apoptosis 유발과 밀접한 관련이 있음을 알 수 있었다. RT-PCR 및 Western blot 분석 결과, sulforaphane 처리에 의하여 cyclin A 및 cyclin B1, Cdc2의 발현이 단백질 수준에서 선택적으로 저하되었으며, 종양억제 유전자 p53 및 Cdk inhibitor p21의 발현은 전사 및 번역 수준에서 sulforaphane 처리 농도 의존적으로 증가되었다. Sulforaphane의 항암 기전을 규명하기 위해서는 더 많은 연구가 부가적으로 필요하겠지만, 본 연구의 결과들에 의하면 sulforaphane은 강력한 인체암세포의 증식 억제 및 항암작용이 있을 것을 시사하여 준다고 할 수 있다.

• Archives of Pharmacal Research
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• 제28권1호
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• pp.73-80
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• 2005

The effect of capsaicin on apoptotic cell death was investigated in HepG2 human hepatoma cells. Capsaicin induced apoptosis in time- and dose-dependent manners. Capsaicin induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration, and BAPTA, an intracellular $Ca^{2+}$ chelator, significantly inhibited capsaicin-induced apoptosis. The capsaicin-induced increase in the intracellular $Ca^{2+}$ and apoptosis were not significantly affected by the extracellular $Ca^{2+}$ chelation with EGTA, whereas blockers of intracellular $Ca^{2+}$ release (dantrolene) and phospholipase C inhibitors, U-73122 and manoalide, profoundly reduced the capsaicin effects. Interestingly, treatment with the vanilloid receptor antagonist, capsazepine, did not inhibit either the increased capsaicin-induced $Ca^{2+}$ or apoptosis. Collectively, these results suggest that the capsaicin-induced apoptosis in the HepG2 cells may result from the activation of a PLC-dependent intracellular $Ca^{2+}$ release pathway, and it is further suggested that capsaicin may be valuable for the therapeutic intervention of human hepatomas.

• Archives of Pharmacal Research
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• 제28권10호
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• pp.1183-1189
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• 2005

Although plant-derived phenolic acids have been reported to have anti-cancer activity, the exact mechanism is not completely understood. In this study, we investigated the role for reactive oxygen species (ROS) as a mediator of the apoptosis induced by caffeic acid (CA) and ferulic acid (FA), common phenolic acids in plants in HepG2 human hepatoma cells. CA and FA reduced cell viability, and induced apoptotic cell death in a dose-dependent manner. In addition, they evoked a dose-related elevation of intracellular ROS. Treatment with various inhibitors of NADPH oxidase (diphenylene iodonium, apocynin, neopterine) significantly blunted both the generation of ROS and the induction of apoptosis induced CA and FA. These results suggest that ROS generated through activation of NADPH oxidase may play an essential role in the apoptosis induced by CA and FA in HepG2 cells. These results further suggest that CA and FA may be valuable for the therapeutic management of human hepatomas.

• 생명과학회지
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• 제19권9호
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• pp.1251-1257
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• 2009

본 연구에서는 오징어내장을 각 용매별로 분획하여 암세포 성장 억제 효과와 quinone reductase (QR) 유도 활성 증가 효과를 알아보았다. 간암 세포인 HepG2, 유방암 세포주인 MCF-7 그리고 대장암 세포주인 HT-29를 이용하여 암세포 성장 억제 효과를 실험한 결과 모든 세포주에서 TPMH층에서 가장 높은 암세포 성장 억제 효과를 나타내었고 다음으로 TPMM층에서도 높은 암세포 성장 억제 효과를 나타내었다. 그리고 HepG2, MCF-7 및 HT-29 세포주에서 모두 농도 의존적인 암세포 성장 억제 효과를 나타내었으나 간암 세포주인 HepG2에서 가장 높은 효과를 나타내어 특히 간암에 대한 예방효과가 기대되어진다. 또한 암 예방 효과를 알아보기 위하여 3종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 유도 활성 증가 효과를 측정한 결과, 암세포 성장 억제 효과의 결과에서와는 달리 TPMM층에서 유의적으로 가장 높은 QR 유도 활성 증가 효과를 나타내었고 다음으로 TPMH층에서도 높은 QR 유도 활성 증가 효과를 나타내었다. 본 실험 결과에서 암세포 성장 억제 효과는 오징어내장의 hexane 분획층인 TPMH층에서 가장 높게 나타났으며, QR 유도 활성 효과는 methanol 분획층인 TPMM층에서 가장 높게 나타났으므로 이들 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성 추정되어진다. 따라서 본 연구를 통하여 오징어 부산물인 내장을 이용한 항암 관련 기능성 식품의 개발 가능성이 보이며, 이를 위한 TPMH 분획층과 TPMM 분획층에 대한 집중적인 연구가 요구된다.

• Toxicological Research
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• 제19권3호
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• pp.235-240
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• 2003

The modification of CYP2E1 activity is of considerable interest because of its role in the metabolic activation of a variety of toxic chemicals. In the present studies, the time-course of changes in human CYP2E1 activities was determined after treatment with ethanol, glycerol, 4-methylpyrazole or isoniazid using intact HepG2 cells transfected by human CYP2E1. Hydroxylation of chlorzoxazone was chosen for the measurement of CYP2E1 activity. CYP2E1 protein levels were increased upon cultivation of cells in the presence of ethanol, glycerol, 4-methylpyrazole or isoniazid for 24 hr. After 24 hr cultivation, ethanol or glycerol increased CYP2E1 activities, whereas 4-methylpyrazole or isoniazid inhibited. This different effect of the chemical inducers on CYP2E1 activi-ties persisted to subsequent 24 hr. Competitive inhibition study suggested that 4-methylpyrazole or isoniazid has stronger binding affinity to CYP2E1 than ethanol or glycerol. These results demonstrate that different binding affinity of the chemical inducers to the active site of CYP2E1 plays important role in determining real CYP2E1 activity in intact cells after treatment with the chemical inducers. Present study would be helpful in precise understanding of human CYP2E1-mediated toxicity.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.88.2-88.2
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• 2003

Ascorbic acid (vitamin C) has been shown to have anti-cancer actions. However, the exact mechanism of this action is not fully understood. In this study we investigated the possible mechanism of anti-cancer action of ascorbic acid in HepG2 human hepatoblastoma cells. Ascorbic acid induced apoptotic cell death in a dose-dependent manner in the HepG2 cells, assessed by the flow cytometric analysis of hypodiploid nuclei stained with propodium iodide. In addition, ascorbic acid increased intracellular Ca$\^$2+/ concentration, whereas the level of reactive oxygen species was not significantly changed, suggesting that ascorbic acid may not alter cellular redox potential in the cells. (omitted)

• Archives of Pharmacal Research
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• 제27권3호
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• pp.305-313
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• 2004

The effect of diazoxide, a $K^{+}$channel opener, on apoptotic cell death was investigated in HepG2 human hepatoblastoma cells. Diazoxide induced apoptosis in a dose-dependent manner and this was evaluated by flow cytometric assays of annexin-V binding and hypodiploid nuclei stained with propidium iodide. Diazoxide did not alter intracellular $K^{+}$concentration, and various inhibitors of $K^{+}$channels had no influence on the diazoxide-induced apoptosis; this implies that $K^{+}$channels activated by diazoxide may be absent in the HepG2 cells. However, diazoxide induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration, and this was completely inhibited by the extracellular $Ca^{2+}$ chelation with EGTA, but not by blockers of intracellular $Ca^{2+}$ release (dantrolene and TMB-8). This result indicated that the diazoxide-induced increase of intracellular $Ca^{2+}$ might be due to the activation of a Ca2+ influx pathway. Diazoxide-induced $Ca^{2+}$ influx was not significantly inhibited by either voltage-operative $Ca^{2+}$ channel blockers (nifedipinen or verapamil), or by inhibitors of $Na^{+}$, $Ca^{2+}$-exchanger (bepridil and benzamil), but it was inhibited by flufenamic acid (FA), a $Ca^{2+}$-permeable nonselective cation channel blocker. A quantitative analysis of apoptosis by flow cytometry revealed that a treatment with either FA or BAPTA, an intracellular $Ca^{2+}$ chelator, significantly inhibited the diazoxide-induced apoptosis. Taken together, these results suggest that the observed diazoxide-induced apoptosis in the HepG2 cells may result from a $Ca^{2+}$ influx through the activation of $Ca^{2+}$-permeable non-selective cation channels. These results are very significant, and they lead us to further suggest that diazoxide may be valuable for the therapeutic intervention of human hepatomas.

• 한국식품영양과학회지
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• 제34권1호
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• pp.8-12
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• 2005

본 연구는 약용 및 식용 식물로 사용되고 있는 쑥부쟁이 추출물과 분획물의 암세포 증식 억제 효과와 QR유도 효과를 실험하였다. 쑥부쟁이의 metanol 추출 및 용매 분획물의 암세포 증식억제 효과를 MTT assay로 실험한 결과, 3종의 인체 암세포주, 간암세포주인 HepG2, 자궁경부암 세포주인 HeLa 및 유방암 세포주인 MCF-7에서 모두 쑥부쟁이의 ethyl ether 분획층인 AYMEE층과 ethyl acetate 분획층인 AVMEA층에서 암세포 성장 저지 효과가 제일 컸다. 특히 HepG2 세포주에서 는 AYMEE를 200 $\mu$g/mL 첨가시켰을 때 94.5%의 암세포 증식 억제 효과가 나타났으며, 농도를 증가시킬수록 그 효과가 커져 500 $\mu$g/mL에서는 97.7%의 저지 효과를 보였다. MCF-7 세포주와 HeLa 세포주에서도 이와 비슷한 경향이었다. Butanol층인 AYMB와 메탄올층인 AYM에서는 대체적으로 낮은 농도에서는 효과가 약하였으나 300 $\mu$g/mL 이상에서는 높은 암세포 성장저지 효과를 보였다. HepG2 세포를 이용하여 암예방 지수인 QR 유도 활성을 측정한 결과, 다른 분획층에 비해 비극성 용매층인 hexane 분획층, AYMH에서 QR 유도 활성을 증가시키는 경향이었다.