• 한국식품위생안전성학회지
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• 제28권2호
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• pp.146-151
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• 2013

t-BHP는 대표적인 산화스트레스에 의한 간 손상 모델이며, 본 연구자는 랫드에 기능성이 강화된 기능성 들깻잎 추출물(P. frutescens leaf extract, PLE)을 250, 500 또는 1000 mg/kg body weight (b.w.)으로 6일간 경구 투여하고 t-BHP의 복강 주사로 간 손상을 일으킨 후 기능성 들깻잎 추출물이 간에서 산화스트레스를 얼마나 억제하여 주는지를 혈액의 간 손상 지표인 lactate dehydrogenase (LDH), aspartate aminotransferase (AST) 그리고 alanine aminotransferase (ALT)를 측정하였으며, 간 조직에서 항산화 바이오 마커인 reduced glutathione (GSH), 지질과산화물의 척도인 malondialdehyde (MDA)를 통해서 측정하였다. 산화스트레스에 의한 간 손상시 GSH는 아무것도 처리하지 않은 정상 대조군($146.0{\pm}5.4$ mM GSH/g protein)에 비해 t-BHP로 산화스트레스를 유발한 그룹에서 $128.6{\pm}6.8$ mM GSH/g protein로 감소하였다. 반면 기능성 들깻잎 추출물을 250, 500 그리고 1000 mg/kg b.w. 을 투여한 그룹에서는 $129.3{\pm}2.6$ mM GSH/g protein, $151.9{\pm}6.8$ mM GSH/g protein, $171.9{\pm}5.2$ mM GSH/g protein로 농도 의존적으로 GSH 함량이 회복되는 경향을 나타내며, 500 mg/kg b.w. 투여군부터 정상 대조군의 GSH 함량과 비슷한 수준을 나타내었다. 또한, 간 조직에서 산화스트레스에 의하여 발생된 지질과산화물을 측정하였을 때 t-BHP에 의하여 산화스트레스만 유발한 그룹은 $834.0{\pm}154.7{\mu}M/g$ protein 로 정상 대조군의 $385.6{\pm}39.7{\mu}M/g$ protein 보다 2.17배 높은 MDA를 생성한 것으로 지질과산화가 많이 일어난 것을 확인하였으나, 기능성 들깻잎 추출물을 투여한 그룹의 MDA의 생성량은 $669.2{\pm}145.0$, $595.1{\pm}142.6$, $415.9{\pm}133.8{\mu}M/g$ protein 로 농도에 따라 감소하는 경향을 확인하였다. 랫드의 간 조직에서, 독성을 유발하는 t-BHP를 복강 투여 후 기능성 들깻잎 추출물을 경구 투여하였을 때 지질과산화의 지표인 MDA의 감소와 항산화의 지표인 GSH 함량이 증가하였다. 조직병리학적 확인을 위하여 간 조직을 H&E 염색 후 광학현미경으로 관찰하였을 때, t-BHP만 처리한 음성 대조군의 경우 간 세포의 괴사와 조직의 변형을 확인할 수 있었지만 기능성 들깻잎 추출물을 처리하였을 경우 모두 정상 상태의 조직을 관찰할 수 있었다. 위의 결과들을 종합하였을 때, t-BHP가 간에서 산화스트레스를 유발하여 간 손상을 야기시키고, 간 조직의 인지질 막 손상을 줄 수 있으며, 기능성 들깻잎 추출물은 간 손상에 대한 보호 효과가 있음을 확인하였다.

• 생약학회지
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• 제38권3호통권150호
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• pp.287-290
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• 2007

Phytochemical investigation of the MeOH extract of the dried stem barks of Fraxinus rhynchophylla Hance (Oleaceae), as guided by cytoprotective activity against tert-butyl hydroperoxide (t-BHP)-induced cell injury in mouse hippocampal HT22 cells, furnished two coumarins, esculetin (1) and fraxetin (2). Compounds 1 and 2 had the significant cytoprotective effects on t-BHP-induced cellular oxidative injury in HT22 cells. Furthermore, compounds 1 and 2 showed potent DPPH radical scavenging effect, exhibiting $IC_{50}$ values of 14.68 and 9.64 ${\mu}M$, respectively.

• Natural Product Sciences
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• 제12권3호
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• pp.134-137
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• 2006

Four lignan compounds including gomisin J (1), schizandrin (2), gomisin A (3), and angeloyl gomisin H (4) have been isolated from the MeOH extract of Schizandra chinensis fruits. The evaluation for protective effect of compounds 1-4 against tert-butyl hydroperoxide (t-BHP)-induced cytotoxicity in hippocampal HT22 cell line was conducted. Compound 1 showed significant protective effect with an $EC_{50}$ value of $43.3{\pm}2.3\;{\mu}M$, whereas compounds 2-4 were inactive. Trolox, one of the well-known antioxidant, used as a positive control, and also showed protective effect with an $EC_{50}$ value of $213.8{\pm}8.4\;{\mu}M$. These results suggest that compound 1 may possess the neuroprotective activity against oxidant-induced cellular injuries.

• 대한약침학회지
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• 제10권1호통권22호
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• pp.121-135
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• 2007

Objectives : This study was carried out to examine protective effect of wild ginseng extract on HepG2 human hepatoma cell line against tert-Butyl hydroperoxide (t-BHP)-induced oxidative damage. Methods : To evaluate protective effect of wild ginseng extract against t-BHP induced cytotoxicity, LDH level and activity of glutathione peroxidase and reductase were measured. Gene expression was also measured using DNA microarray. Results : Wild ginseng extract showed a significant protective effect against t-BHP-induced cytotoxicity in HepG2 cell line. It is not, however, related with the activities of glutathione peroxidase and glutathione reductase. Analysis of gene expression using DNA chip, demonstrated that 28 genes were up-regulated in t-BHP only group. Five genes - selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, serfiredoxin 1 homolog - may be related with the protective effect of wild ginseng extract. Conclusions : Based on the results, a protective effect of wild ginseng extract against t-BHP-induced oxidative damage in HepG2 cell line is not associated with the activities of glutathione peroxidase and glutathione reductase, but with the expression of selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, and serfiredoxin 1 homolog.

• Food Science and Biotechnology
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• 제17권5호
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• pp.1086-1091
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• 2008

The antioxidant activities of the extracts of dangyuja (Citrus grandis Osbeck) leaves were evaluated. The highest phenolic content was obtained from the ethyl acetate fraction (EF) (202.1$\pm$0.8 mg GAE/g dried extract) and it exhibited the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. The cytoprotective effects of EF on oxidative damage induced by tert-butyl hydroperoxide (t-BHP) in a human hepatoma cell line, HepG2 cells, were investigated to understand the intracellular antioxidant mechanisms. Treatment of HepG2 cells with EF prior to oxidative stress was found to inhibit reactive oxygen species (ROS) generation, lipid peroxidation, and DNA damage in a dose-dependent manner. Gas chromatography-mass spectrometry (GC-MS) studies on EF resulted in tentative identification of 19 compounds representing 94.3% of the total content. Taken together, these results demonstrated that EF has excellent antioxidant activities and thus dangyuja leaves have great potential as a source for natural antioxidant which can be applied in food products.

• 생명과학회지
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• 제31권3호
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• pp.338-345
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• 2021

고장초는 오랫동안 식용 및 의약 목적으로 차를 만드는 데 사용되어 왔다. 그러나 고장초를 고부가가치 식용 재료로 사용하는 연구는 불충분하다. 이전 연구에서 우리는 tricin이 고장초의 주요 화합물임을 확인했다. 본 연구에서는 고장초 추출물의 HepG2 세포 보호 효과를 조사하였다. 고장초 추출물 또는 tricin의 HepG2 세포에 대한 독성 실험 결과, 사용한 모든 농도에서 고장초 추출물 또는 tricin에 대한 독성은 나타나지 않았다. 또한 tert-butyl hydroperoxide (t-BHP)에 의해 감소된 세포 생존율은 고장초 추출물(100 ㎍/ml에서 69.23%) 또는 tricin (100 ng/ml에서 74.47%)에 의해 증가되었다. 또한 고장초 추출물(250 ㎍/ml) 또는 tricin (250 ng/ml)은 산화적 손상에 의한 활성 산소종(ROS) 생성을 효과적으로 억제했다. 또한 항산화 효소로 알려진 superoxide dismutase 1(SOD1), catalase, heme oxygenase-1 (HO-1), nuclear factor erythroid-related factor 2 (Nrf2)의 단백질 발현은 고장초 추출물(100 ㎍/ml에서 2.1, 1.6, 1.7, 1.5배) 또는 tricin (250 ng/ml에서 2.6, 1.8, 2.2, 2.0배) 처리로 증가했으며, 이는 고장초 추출물의 HepG2 세포 보호 효과는 tricin의 활성과 관련이 있다는 것을 시사한다. 종합하면 고장초는 간 기능개선 관련 기능성 식품 소재로 개발이 가능할 것으로 기대된다.

• 생명과학회지
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• 제10권5호
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• pp.475-483
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• 2000

Normally, aerobic cells are protected from the damage of free radicals by antioxidative enzymes such as catalase, superoxide dismutase (SOD), glutathione (GSH) peroxidase and GSH-S-transferase. In this study, we have investigate the effect of egg yolk protein hydrolysates on antioxidative activity and the activity of antioxidative enzyme in cultured hepatocytes (Chang). Without the pretreatment with hydrolysate, about 50% of the hepatocytes were killed within 2h by 225$\mu$M tert-butyl hydroperoxide (t-BHP). By contrast, fewer than 20% of the 5 K hydrolysate (permeate from 5 kDa membrane and not passed through 1 kDa membrane)-pretreated hepatocytes were killed by the same concentrations of t-BHP. In addition, the activities of catalase, GSH peroxidase and GSH-transferase were significantly increasing with the treatment of 5 K hydrolysate. These results suggest that 5 K hydrolysate exerts antioxidative effect by increasing activity of antioxidative enzymes.

• 대한의생명과학회지
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• 제20권2호
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• pp.56-61
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• 2014

This study attempted to confirm the antioxidative potential of luteolin against tert-butyl hydroperoxide (t-BHP) induced oxidative damage and to investigate its molecular mechanism related to glutathione (GSH)-dependent enzymes in HepG2 cells. Treatment with luteolin resulted in attenuation of t-BHP induced generation of reactive oxygen species (ROS) and oxidative stress-mediated cell death. In addition, accelerated expression of GSH-dependent antioxidative enzymes, glutathione peroxidase (GPx) and glutathione reductase (GR), and heme oxygenase (HO)-1, as well as strengthened GSH content was induced by treatment with luteolin, which was in accordance with increased nuclear translocation of nuclear factor-erythroid 2 p45-related factor 2 (Nrf2), a transcription factor for phase 2 enzymes, in a dose-dependent manner. These results suggest that the cytoprotective potential of luteolin against oxidative damage can be attributed to fortified GSH-mediated antioxidative pathway and HO-1 expression through regulation of Nrf2 in HepG2 cells.

• 대한한의학방제학회지
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• 제31권4호
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• pp.231-243
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• 2023

Objectives : Jasminum officinale L. var. grandiflorum is used as a traditional or folk remedy in China to treat arthritis, hepatitis, duodenitis, conjunctivitis, gastritis, and diarrhea. In this study, we aimed to study the hepatocyte protective activity and molecular mechanism of Jasminum officinale L. var. grandiflorum aqueous extract (JGW) using HepG2 hepatocyte cell lines. Methods : HepG2 cells were pretreated with diverse concentrations of JGW, and then the cells were exposed to tert-butyl hydroperoxide (tBHP) for inducing oxidative stress. Hydrogen peroxide (H₂O₂) production, glutathione (GSH) concentration, mitochondrial membrane potential (MMP) and cell viability were measured to investigate hepato-protective effects of JGW. Phosphorylation of AMP-activated protein kinases (AMPK), acetyl coenzyme A carboxylase (ACC) and effects of compound C on cell viability were examined to observe the role of AMPK on JGW-mediated cytoprotection. Results : Pretreatment with JGW (10-300 ㎍/mL) significantly suppressed cytotoxicity induced by tBHP in a concentration dependent manner and reduced the expression of cleaved PARP and cleaved caspase-3 proteins related to apoptosis in HepG2 cells. In addition, pretreatment with JGW significantly prevented the increase in H₂O₂ production, GSH depletion, and lower MMP induced by tBHP. Treatment with JGW (30 minutes of incubation and concentrations of 100 and 300 ㎍/mL) increased the phosphorylation of AMPK and ACC and treatment with compound C, a chemical inhibitor of AMPK, inhibited the cytoprotective effect of JGW. Conclusions : Our results demonstrated that JGW may protect hepatocytes from oxidative stress via activation of AMPK.

• Natural Product Sciences
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• 제17권4호
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• pp.285-290
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• 2011

The rhizome of Alisma orientale Juzep (Alismataceae) has been used as a crude drug for diabetes, edema, inflammation and urinary disturbances in oriental medicine. Recent animal studies have shown that the extract of Alisma orientale rhizome (AOR) can potently lower high levels of serum lipids and improve insulin resistance, which are usually detected in patients and animals with non-alcoholic fatty liver disease. So, we studied the antioxidative effects of AOR extracts and fraction in vitro and their protective effects against acute hepatotoxicity induced by $CCl_4$ in vivo.. We then investigated the effects of each fraction on hepatotoxicity induced by tert-butyl hydroperoxide (t-BHP). DAOR (dichloromethane fraction of the Alisma orientale rhizome) scavenged free radicals and superoxide anions. DAOR protected against $CCl_4$ induced hepatotoxicity. DAOR had hepatoprotective and antioxidative effects against t-BHP-induced hepatotoxicity in HepG2 cells and in rats.