• Biomolecules & Therapeutics
• /
• 제12권3호
• /
• pp.189-193
• /
• 2004

Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

• Natural Product Sciences
• /
• 제25권3호
• /
• pp.228-232
• /
• 2019

Five secondary metabolites, including a new isopimarane derivative xylaroisopimaranin A (1), were isolated from the endophytic fungus Xylaralyce sp. (HM-1), and their structures were elucidated by 1D, 2D NMR, MS and CD spectra. Their bioactivities were performed to antibacterial, Hep G2 cells cytotoxicity and brine shrimp inhibition. The biological evaluation results showed that the xylaroisopimaranin A (1), xylabisboein B (2), griseofulvin (3), 5-methylmellein (4) and mellein-5-carboxlic acid (5) displayed no significant Hep G2 cells cytotoxicity and antibacterial acitivity, but they inhibited the brine shrimp with $IC_{50}$ from 0.5 to $25{\mu}mol/mL$.

• 대한한의학방제학회지
• /
• 제30권3호
• /
• pp.155-163
• /
• 2022

Objectives : Lonicera japonica is known for anti-inflammation and antibiotic effect in Korean medicine. This study aimed for investigating the cytoprotective effect of Lonicera japonica extract (LJE) for HepG2 cells against arachidonic acid (AA)+iron-induced oxidative stress. Methods : The effect of LJE on cell viability was assessed by MTT assay. ROS assay was selected to assess antioxidant effect of LJE. To assess LJE's effect on mitochondrial function, flow cytometric analysis was operated. And immunoblot analysis was used to establish the underlying mechanism of LJE. Results : LJE protected HepG2 cells against AA+iron-induced oxidative stress by phosphorylation of liver kinase B1 and blocked the decline of procaspase 3. Also, LJE preserved the mitochondrial membrane permeability induced by AA+iron. Conclusion : LJE protected the hepatocyte from AA+iron-induced oxidative stress by activation of LKB1 by the preservation of mitochondrial functions.

• 대한한방내과학회지
• /
• 제18권1호
• /
• pp.175-190
• /
• 1997

In order to investigate the effects of Gamihagochosan Extract(加味夏枯草散抽出液) on antitumor effects after human cell lines (A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurement of antitumor effect of Mitomycin C(MMC) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for the measurement of antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows : 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 Cells after exposure to the extract of Gamihagochosan extract depressed the growth of tumor cells by concentration of Garnihagochosan. 2. Antitumor activity of the ethanol extract from Gamihggochosan extract and MMC on ascites form of Ehrlich carcinoma in mice is slightly improved. Especially the mean of survival times in the group of 200mg/kg and MMC 0.1mg/kg is improved over 34.9%. 3. When Gamihggochosan extract and MMC are administered together, the weight of tumor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Gamihagochosan extract and MMC are more significantly improved than MMC alone. According to the above result, it could be suggested that Gamihagochosan extract has indirect antitumor effect by the increase of MMC uptake.

• 대한한의학회지
• /
• 제18권2호
• /
• pp.119-136
• /
• 1997

In order to investigate the effects of Soyosangamibang Extract(逍遙散加味方抽出液) on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 cells after exposure to the extract of Soyosangamibang extract depressed the growth of tumor cells by concentration of Soyosangamibang, 2. Antitumor activity of the ethanol extract from Soyosangamibang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of 200mg/kg and MMC 0.1mg/kg is improved Over 50%. 3. WhenSoyosangamibang extract and MMC are administrated together, the weight of turnor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Soyosangarmibang extract and MMC are more significantly improved than MMC alone. According to the above results, it could be suggested that Soyosangamibang extract has indirect antitumor effect by strengthen the effect of MMC.

• 한국약용작물학회지
• /
• 제10권2호
• /
• pp.109-115
• /
• 2002

황칠나무 잎의 추출물의 면역활성증진 실험에서 인간정상 간세포의 경우 모든 추출물이 1.0 Mg/ml의 농도에서 최고 26%이하의 세포독성을 나타내었다. 서로 다른 4가지의 암세포주(MCF7, A549, Hep3B, AGS)에서 50%이상의 저해율을 나타냈고, 정상 세포의 결과와 암세포의 저해율을 비로 나타낸 selectivity의 측정에서 모든 암세포주가 1.5이상의 사멸도를 나타내었고 전체적으로 에탄올 추출물의 효과가 가장 좋았다. 에탄올 추출물의 경우에서 인감 유방암 세포주(MCF7)와 인간 간암세포주(Hep3B)의 경우에서 1.0 mg/ml농도에서 각각 65%와 67%의 저해율을 기록했다. 면역세포 실험에서 에탄올 추출물이 1.0 mg/ml의 농도에서 B세포는 1.22배, T세포는 1.27배의 촉진 활성을 보였고, 6일 동안측정한 $cytokines(IL-6,\;TNF-{\alpha})$의 양도 에탄올 추출물의 경우 T cell의 경우 IL-6은 94pg/m1, $TNF-{\alpha}$은 75pg/ml로 증가하는 것을 알 수 있었다. 이상의 결과로 볼 때 추출 열과 추출용매 등에 의해 황칠나무 잎에 존재하는 여러 가지 유용성분들이 풍부하게 존재하며, crude추출물 중에 에탄올 추출물이 면역활성에서 좋은 효과를 보였다는 것을 알 수가 있었고, 이를 통해서 황칠나무 잎을 이용한 기능성 식품으로의 연구 개발이 통해서 충분히 그 가치가 있다는 것을 알 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제22권1호
• /
• pp.156-158
• /
• 2012

A synthetic coprisin analog peptide, 9-mer dimer CopA3 (CopA3) was designed based on a defensin-like peptide, Coprisin, isolated from the bacteria-immunized dung beetle Copris tripartitus. Here, CopA3 was investigated for its antimicrobial activity and cancer cell growth inhibition. CopA3 showed antimicrobial activities against various pathogenic bacteria and yeast fungus with MIC values in 2~32 ${\mu}M$ ranges, and inhibited the cell viabilities of pancreatic and hepatocellular cancer cells, except MIA-Paca2, Hep3B, and HepG2 cells, in a dose-dependent manner. The average $IC_{50}$ values of CopA3 against pancreatic and hepatocellular cancer cells were 61.7 ${\mu}M$ and 67.8 ${\mu}M$, respectively. The results indicate that CopA3 has potential in the treatments of pancreatic and hepatocellular cancers as well as microorganism infection disease.

• Journal of Ginseng Research
• /
• 제43권3호
• /
• pp.452-459
• /
• 2019

Background: Ginsenoside compound K(C-K), a major metabolite of ginsenoside, exhibits anticancer activity in various cancer cells and animal models. A cell signaling study has shown that C-K inhibited nuclear factor-kappa B ($NF-{\kappa}B$) pathway in human astroglial cells and liver cancer cells. However, the molecular targets of C-K and the initiating events were not elucidated. Methods: Interaction between C-K and Annexin A2 was determined by molecular docking and thermal shift assay. HepG2 cells were treated with C-K, followed by a luciferase reporter assay for $NF-{\kappa}B$, immunofluorescence imaging for the subcellular localization of Annexin A2 and $NF-{\kappa}B$ p50 subunit, coimmunoprecipitation of Annexin A2 and $NF-{\kappa}B$ p50 subunit, and both cell viability assay and plate clone formation assay to determine the cell viability. Results: Both molecular docking and thermal shift assay positively confirmed the interaction between Annexin A2 and C-K. This interaction prevented the interaction between Annexin A2 and $NF-{\kappa}B$ p50 subunit and their nuclear colocalization, which attenuated the activation of $NF-{\kappa}B$ and the expression of its downstream genes, followed by the activation of caspase 9 and 3. In addition, the overexpression of Annexin A2-K320A, a C-K binding-deficient mutant of Annexin A2, rendered cells to resist C-K treatment, indicating that C-K exerts its cytotoxic activity mainly by targeting Annexin A2. Conclusion: This study for the first time revealed a cellular target of C-K and the molecular mechanism for its anticancer activity.

• 한국식품영양과학회지
• /
• 제35권10호
• /
• pp.1297-1303
• /
• 2006

본 연구는 쌀겨의 사람유래 암세포 증식 억제효과(in vitro)와 사람유래 비만세포주(HMC-1)의 활성 억제효과 및 흰쥐 복강 비만세포로부터 히스타민 분비에 미치는 영향을 관찰하였다. Hep3B와 HeLa 세포에 대한 형태변화는 쌀겨의 물 추출물과 60% 에탄을 추출물 처리군 모두 농도에 비례하여 암세포의 밀도가 감소하는 경향을 보였고, 시료를 고농도로 처리할수록 사멸세포 형태인 원형의 세포 수가 증가하였다. Hep3B 세포에 대한 증식 억제능은 쌀겨 물 추출물이 60% 에탄을 추출물보다 높았고, $100{\mu}g/mL$와 $1,000{\mu}g/mL$ 농도에서 물 추출물이 60% 에탄을 추출물보다 각각 20%, 55% 더 높은 효과가 있었다. HeLa세포에 대한 증식 억제능은 쌀겨 물 추출물과 60% 에탄을 추출물에서 비슷한 경향을 보였으나 $1,000{\mu}g/mL$와 $3,000{\mu}g/mL$ 농도에서 물 추출물이 60% 에탄을 추출물보다 각각 5%, 27% 더 높았다. 사람의 비만세포주(HMC-1)에서 $Fc{\varepsilon}RI$, tryptase, c-kit mRNA 발현양에 미치는 효과를 관찰한 결과 GAPDH의 발현양은 대조군과 시료를 처리한 실험군에서 모두 비슷하게 나타났다. 비만세포의 $Fc{\varepsilon}RI$ mRNA와 c-kit의 mRNA 발현양은 쌀겨 물 추출물군보다 60% 에탄올 추출물에서 더 많이 억제되는 것으로 나타났으나 tryptase mRNA 발현양은 억제하지 못한 것으로 관찰되었다. 비만세포의 히스타민 분비에 미치는 영향은 무처리 대조군에 비해 $5{\mu}g/mL$ compound 48/80을 처리하였을 때 히스타민 분비량이 유의하게 증가하였으며, 쌀겨 물 추출물 $0.01{\mu}g/mL$에서 약 87%의 억제율을 나타내었고, 에탄을 추출물 $100{\mu}g/mL$에서 약 86%의 억제율을 보였다.

• 한국식품저장유통학회지
• /
• 제15권2호
• /
• pp.274-279
• /
• 2008

해양심층수 간장 및 일반간장을 제조하여 항돌연변이원성과 세포독성을 측정하였으며 sarcoma-180 cell을 이용하여 in vivo에서 항암효과를 살펴보았다. S. typhimurium TA98과TA100 균주를 이용한 실험에서 모든 시료에서 돌연변이원성이 없었으며, 항돌연변이원성 실험에서는 직접변이원인 MNNG($0.4{\mu}g$/plate)의 경우 TA100 균주에서 해양심층수간장의 시료농도 $200{\mu}g$/plate에서 90.9%의 높은 억제효과를 나타내었으며 4NQO($0.15{\mu}g$/plate)에 대해서는 같은 시료농도에서 62.0%의 억제효과를 나타내었다. 그리고 4NQO의 경우 TA98 균주에 대해서 해양심층수 간장은 61.7%의 억제효과를 나타내었으며 모든 시료는 농도 의존적으로 억제하는 것으로 나타났다. 세포독성 효과를 알아보기 위하여 HeLa, Hep3B, AGS, A549와 MCF-7을 사용하였다. 각 시료 추출물의 암세포 성장효과를 조사한 결과, 해양심층수 간장이 1 mg/mL의 농도에서 각각 69.4%, 70.5%, 55.6%, 82.1% 및 73.2%의 억제율을 나타내었다. 그리고 해양심층수 간장은 고형암 성장 억제 실험에서 대조군에 비해서 40.9%의 고형암 성장 억제효과를 나타내었다.