• 생명과학회지
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• 제24권6호
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• pp.595-602
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• 2014

미더덕껍질(Styela clava tunic, SCT)은 항염증 복합체, 창상필름, 골재생 유도 등을 포함한 다양한 의학적인 치료영역에 이용 되고 있다. 본 연구에서는 미더덕껍질 열수추출물(aqueous extract of Styela clava tunic, AE-SCT)의 $H_2O_2$에 의해 유발된 세포사멸의 보호 효과를 알아보기 위하여 세포 활성도의 변화에 관련된 요인을 측정하였다. 그 결과, AE-SCT는 3.3 mg/g의 플라보노이드와 32.3 mg/g의 페놀화합물을 포함하고 있었으며, HepG2 세포주에 독성을 유발하지 않았다. 또한, $H_2O_2$ 처리 후 AE-SCT를 처리하는 실험에서 AE-SCT는 $H_2O_2$에 의해 유발된 세포사멸을 개선하는 효과를 나타내지 않았다. 그러나, $H_2O_2$ 처리전에 AE-SCT를 사전 처리하는 예방효과 실험에서, 세포생존율은 $H_2O_2$만 처리한 그룹에 비하여 AE-SCT를 처리한 그룹에서 유의적으로 증가하였으며, 특히 AE-SCT를 $50{\mu}g/ml$ 처리한 농도에서 가장 높았다. 또한, FACS분석과 DAPI 염색에서도 사멸 세포의 수는 $H_2O_2$만 처리한 그룹에 비하여 AE-SCT를 처리한 그룹에서 유의적으로 감소하였다. 더불어, $H_2O_2$의 처리에 의해 유도된 Bax/Bcl-2 발현비율은 AE-SCT처리에 의해 농도의존적으로 감소되었다. 이러한 연구 결과는 AE-SCT가 $H_2O_2$에 의해 유발된 세포사멸을 예방하는 우수한 효과를 가지고 있음을 제시하고 있어 향후 다양한 항산화 제품 개발을 위한 기초자료로 사용될 수 있을 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제24권5호
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• pp.373-384
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• 2020

Paeonol, quercetin, β-sitosterol, and gallic acid extracted from Moutan Cortex had been reported to possess anti-oxidative, anti-inflammatory, and anti-tumor activities. This work aimed to illustrate the potential anti-oxidative mechanism of monomers in human liver hepatocellular carcinoma (HepG2) cells-induced by hydrogen peroxide (H₂O₂) and to evaluate whether the hepatoprotective effect of monomers was independence or synergy in mice stimulated by carbon tetrachloride (CCl₄). Monomers protected against oxidative stress in HepG2 cells in a dose-response manner by inhibiting the generation of reactive oxygen species, increasing total antioxidant capacity, catalase and superoxide dismutase (SOD) activities, and activating the antioxidative pathway of nuclear factor E2-related factor 2/Kelch-like ECH-associated protein 1 (Nrf2/Keap1) signaling pathway. We found that the in vitro antioxidant capacities of paeonol and quercetin were better than those of β-sitosterol and gallic acid. Furthermore, paeonol apparently diminished the levels of alanine transaminase and aspartate aminotransferase, augmented the contents of glutathione and SOD, promoted the expressions of Nrf2 and heme oxygenase-1 proteins in mice stimulated by CCl₄. In HepG2 cells, paeonol, quercetin, β-sitosterol, and gallic acid play a defensive role against H₂O₂-induced oxidative stress through activating Nrf2/Keap1 pathway, indicating that these monomers have anti-oxidative properties. Totally, paeonol and quercetin exerted anti-oxidative and hepatoprotective effects, which is independent rather than synergy.

• 대한한방내과학회지
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• 제29권2호
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• pp.490-499
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• 2008

Objectives : This study was aimed to elucidate the effects of Trichosanthes kirilowii extract (TKE) on the angiogenesis and growth of tumor cells. Methods : Tube formation assay was performed by using human umbilical vein endothelial cells (HUVEC), and anchorage dependent colony assay was performed by using B16-F10 melanoma, Hep G2 and HT1080, CT-26 and SNU-1 cells. Results : For HUVEC, TKE at a level of more than 100 ${\mu}g/m{\ell}$ suppresses cell growth. For HUVEC at 100 ${\mu}g/m{\ell}$ and greater TKE density, the formation of tubes was suppressed in a dose-dependant manner. TKE controls the colony formations of B16-F10 melanoma cells, CT 26 cells, and Hep G2 cells, and its effect is proportional to density. In HT1080 cells and SNU-1 cells, formation is suppressed regardless of density. Conclusions : From these results, it could be concluded that TKE has significant properties on anti-angiogenesis and growth inhibiting of tumor cells. It is suggested that TKE will be a good candidate for new drugs or therapeutics for anti-angiogenesis.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1399-1404
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• 2006

The roots of Aralia continentalis (AC) have been used traditionally in Korean as a folk medicine for anti-inflammation and as an anti-rheumatic. In this study, we report that the ethyl acetate-soluble traction (ACE) of the methanolic extract of AC root inhibited the cell growth of various human cancer cell lines and induced apoptosis of HL-60, human promyelocytic leukemia cells. Its $IC_{50}$ values on growth inhibition were estimated to be $56.3{\mu}g/ml$ on HL-60, $87.2{\mu}g/ml$ on HepG2, $93.2{\mu}g/ml$ on HeLa, $135.5{\mu}g/ml$ on DU-145, and $135.8{\mu}g/ml$ on HT-29 cells. Interestingly, ACE showed no antiproliferative effect on normal lymphocyte cells used as control. Furthermore, nuclear DAPI staining revealed the typical nuclear features of apoptosis in the HL-60 cells exposed to $80{\mu}g/ml$ ACE, and a flow cytometric analysis of the HL-60 cells using propidium iodide showed that the apoptotic cell population increased gradually from 5% at 0 h to 16% at 12 h and 20% at 24 h after treated with $50{\mu}g/ml$ of ACE. TUNEL assay also revealed the apoptotic induction of the HL-60 cells treated with ACE. To obtain further information on the ACE-induced apoptosis, the expression level of certain apoptosis-associated proteins was examined using a Western blot analysis. Treatment of the HL-60 cells with ACE resulted in the activation of caspase-3, and subsequent proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). The above results confirmed that the apoptosis in the HL-60 cells was induced by ACE, and that caspase-3-mediated PARP cleavage was involved in the process.

• 생명과학회지
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• 제33권12호
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• pp.1002-1014
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• 2023

다양한 질병의 발병 및 진행을 예방하기 위해 오랫동안 사용되어 온 대계는 항산화 활성을 포함하여 광범위한 생리 활성을 갖는 것으로 보고되었으나, 간세포에서의 효능에 대한 연구는 여전히 미비한 실정이다. 본 연구에서는 간세포에서 대계 에탄올 추출물(EECJ)의 항산화 활성 및 관련 기전을 조사하기 위해 인간 간세포암종 HepG2 세포주를 선택하였고, 산화적 스트레스를 모방하기 위해 H₂O₂를 처리하였다. 본 연구 결과에 따르면 EECJ는 H₂O₂가 처리된 HepG2 세포에서 세포 생존율의 감소와 LDH의 방출을 유의적으로 억제하였다. EECJ는 또한 세포 형태학적 변화의 관찰, 유세포 분석 및 LC3의 발현 결과에서 입증된 바와 같이 H₂O₂에 의해 유도된 자가포식을 유의하게 감소시켰으며, H₂O₂에 의해 유도된 세포사멸과 세포주기의 교란을 약화시켰다. 이는 H₂O₂에 노출된 HepG2 세포가 EECJ에 의하여 지속적인 세포 분열과 증식이 유지되고 있음을 의미한다. 또한, EECJ가 강력한 항산화 활성을 가지고 있음을 세포 내 ROS 및 미토콘드리아 슈퍼옥사이드 생산의 차단으로 확인하였으며, 이는 H₂O₂에 의해 감소된 세포 내 GSH 함량의 회복과 MnSOD 및 GPx의 발현 및 활성을 보존과 연관성이 있었다. 비록 EECJ에 함유된 활성 성분의 분석과 in vivo 동물 모델에서의 검증이 요구되지만, 본 연구의 결과는 EECJ가 산화적 스트레스로 인한 간세포의 손상을 예방하고 치료하기 위한 잠재적인 후보로 사용될 수 있음을 의미한다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7137-7141
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• 2013

Several studies have shown that nemo-like kinase (NLK) plays a vital role in apoptosis of cancer cells. The present research concerned effects and mechanisms of Taxol on NLK knockdown human laryngeal cancerHep-2 cell lines in vitro. Using RNAi, methyl-thiazoltetrazolium (MTT) assays, real-time RT-PCR, Western blotting and flow cytometry analysis, growth and the cell cycle progression of NLK knockdown Hep-2 cells and expression of downstream molecules were observed. Cell growth was obviously suppressed in the Taxol treated group (P<0.001, 48 hours). Cell numbers of combined Taxol-based chemotherapy with lentivirus mediated RNAi treatment group (Lv-shNLK+Taxol goup) were significantly different from NLK-specific siRNA lentivirus infected group (Lv-shNLK group) (p<0.001). Flow cytometry analysis revealed that Lv-shNLK+Taxol caused the G0/G1-phase DNA content to decrease from 44.1 to 3.33% (p<0.001) and the S-phase DNA content to increase from 38.4 to 82.0% (p<0.001), in comparison with the Lv-shNLK+Taxol group. Immunoblot analysis showed that knockdown of NLK led to significant reduction in the levels of cyclin D1, PCNA and PARP, whereas cyclin B1 was elevated in. Cell growth was also obviously suppressed in the Hep-2 cell line, knockdown of NLK making them more sensitive to Taxol treatment. NLK is expected to become a target of new laryngeal cancer gene therapies.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10439-10444
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• 2015

Many chemotherapeutic agents have been successfully used to treat hepatocellular carcinoma (HCC); however, the development of chemoresistance in liver cancer cells usually results in a relapse and worsening of prognosis. It has been demonstrated that DNA methylation and histone modification play crucial roles in chemotherapy resistance. Currently, extensive research has shown that there is another potential mechanism of gene expression control, which is mediated through the function of short noncoding RNAs, especially for microRNAs (miRNAs), but little is known about their roles in cancer cell drug resistance. In present study, by taking advantage of miRNA effects on the resistance of human hepatocellular carcinoma cells line to cisplatin, it has been demonstrated that miR-340 were significantly downregulated whereas Nrf2 was upregulated in HepG2/CDDP (cisplatin) cells, compared with parental HepG2 cells. Bioinformatics analysis and luciferase assays of Nrf2-3'-untranslated region-based reporter constructor indicated that Nrf2 was the direct target gene of miR-340, miR-340 mimics suppressing Nrf2-dependent antioxidant pathway and enhancing the sensitivity of HepG2/CDDP cells to cisplatin. Interestingly, transfection with miR-340 mimics combined with miR-340 inhibitors reactivated the Nrf2 related pathway and restored the resistance of HepG2/CDDP cells to CDDP. Collectively, the results first suggested that lower expression of miR-340 is involved in the development of CDDP resistance in hepatocellular carcinoma cell line, at least partly due to regulating Nrf2-dependent antioxidant pathway.

• 한국식품과학회지
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• 제40권6호
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• pp.696-701
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• 2008

이 연구의 목적은 더덕 및 도라지 에틸아세테이트 분획물(CLEA나 PGEA)이 다양한 항산화 시스템을 가동시켜 sodium nitroprusside(SNP)에 의해 유도된 산화적 스트레스에 대항하여 세포를 보호해 줄 수 있을지 연구하는 것이다. 0.5 mM의 SNP를 HepG2 세포주에 24시간 동안 노출시켰을 때 세포 생존율이 감소하였으나, CLEA와 PGEA 천처리에 의해 SNP 노출에 의한 세포 생존율 감소가 저해되었다. 또한 항산화 시스템들 발현에 미치는 CLEA와 PGEA의 영향을 RT-PCR로 알아보았다. Catalase, glucose-6-phosphate dehydrogenase(G6PD) 그리고 metallothionein (MT)-1A mRNA 수준이 CLEA를 세포에 24시간 동안 처리한 후 증가하였고, Mn superoxide dismutase, catalase, G6PD, MT-1A 와 MT-2A mRNA 수준이 PGEA 처리에 의해 증가하였다. 우리는 CLEA와 PGEA가 아마 다양한 항산화 시스템들을 증가시키는 간접적인 항산화 효과를 나타내며, 이들 효과들이 산화적 스트레스로부터 세포를 보호해 줄 것이라 추정하였다.

• 한국약용작물학회지
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• 제27권3호
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• pp.208-217
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• 2019

Background: In the present study, we identified two cytotoxic compounds from the root of Rumex crispus L. using a bioassay-based method. Methods and Results: Compared with the other fractions, the diethyl ether ($Et_2O$) fraction of R. crispus root extract exhibited the strongest of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect [scavenging concentration 50% $(SC_{50})=63.8{\pm}1.47{\mu}g/m{\ell}$], nitric oxide (NO) production inhibitory effect on the mouse macrophage cell line RAW264.7 [inhibitory concentration 50% $(IC_{50})=60.9{\pm}7.52{\mu}g/m{\ell}$] and cytotoxicity effect on the human hepatoma cell line, HepG2 [lethal concentration 50% $(LC_{50})=115.4{\pm}1.86{\mu}g/m{\ell}$]. According to the bioassay-based method, two cytotoxic compounds were purified from the $Et_2O$ fraction by using column chromatography and preparative high performance liquid chromatography (prep-HPLC). These two compounds were identified as parietin and chrysophanol by using nuclear magnetic resonance (NMR) and liquid chromatography quadruple time of flight mass spectrometry (LC-QTOF-MS). In addition, both parietin and chrysophanol exhibited a cytotoxicity effect on HepG2 cells, their $LC_{50}$ values were $169.1{\pm}17.67{\mu}M$ and $111.5{\pm}6.62{\mu}M$, respectively. Conclusions: Parietin and chrysophanol isolated from the $Et_2O$ fraction of the R. crispus root extract showed cytotoxicity in HepG2 cell.

• 생약학회지
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• 제38권4호
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• pp.400-402
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• 2007

Tacrine is an acetylcholinesterase inhibitor that is approved for the treatment of Alzheimer's disease. However, tacrine treatment for Alzheimer's disease results in reversible hepatotoxicity in 30-50% of patients, which seriously limits its clinical use. Accordingly, the identification of constituents in natural products that have protective effects on tacrine-induced hepatotoxicity would be valuable. In the present study, an immortalized human hepatoma cell line, HepG2 was employed to screen for agents that protect against tacrine-induced hepatotoxicity. The bioassay-guided fractionation of water extract of Cassiae Semen furnished two anthraquinones, aurantio-obtusin (1) and obtusifolin (2). Compounds 1 and 2 showed hepatoprotective effects with the protection ratio values of 55.3 +/- 0.5% and 41.2 +/- 0.8% at $160{\mu}M$, respectively.