• Asian Pacific Journal of Cancer Prevention
• /
• 제16권5호
• /
• pp.1771-1779
• /
• 2015

Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide. Presently, targeted therapy via monoclonal antibodies to specific tumor-associated antigens is being continuously developed. Hep88 mAb has proven to exert tumoricidal effects on the HepG2 cell via a paraptosis-like morphology. To verify the pathway, we then demonstrated downstream up-regulation of caspase-3, caspase-8 and caspase-9, assessingmRNA expression by real-time PCR and associated enzyme activity by colorimetric assay. Active caspase-3 determination was also accomplished by flow cytometry. Active caspase-3 expression was increased by Hep88 mAb treatment in a dose-and time-dependent manner. All of the results indicated that Hep88 mAb induced programmed cell death in the HepG2 cell line from paraptosis-like to apoptosis by downstream induction of caspases. These conclusions imply that Hep88mAb might be a promising tool for the effective treatment of HCC in the future.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2001년도 추계학술발표대회
• /
• pp.453-456
• /
• 2001

배양된 대황으로 모상근으로부터 추출한 물질에 대한 분획별 신장상피세포에 있어서의 세포 독성을 조사하였다. l 수층과 클로로포름 층으로부터 얻은 대황 모상근 추출물모두 농도의 증가에 따라 세포에 미치는 독성이 증가하였다. 2. 클로로포름층으로부터 얻은 대황 모상근 추출물이 수층으로부터 얻은 대황 추출물보다 세포에 미치는 독성이 크게 나타났다. 콜로로포름층 분획의 $MTT_{50}$, $NR_{50}$, $SRB_{50}$은 각각 289.3 ${\mu}g/ml$. 302.7 ${\mu}g/ml$. 433.8 ${\mu}g/ml$ 이었고, 수층 분획물의 $MTT_{50}$, $NR_{50}$, $SRB_{50}$은 각각 475.8 ${\mu}g/ml$. 428.3 ${\mu}g/ml$. 549.5 ${\mu}g/ml$ 이었다. 3. 세포독성 측정방법에 따라 차이를 보였으며 수층 분획의 경우 NR정량법에서 클로로포름 층 분획의 경우 MTT정량법에서 독성 정도가 더 높게 나타났다.

• 한국식품영양과학회지
• /
• 제31권1호
• /
• pp.131-135
• /
• 2002

본 연구에서는 작약 추출물의 식후 과혈당 억제작용을 조사하였다. 작약 유기용매(hexane, ethyl acetate, butanol, water) 추출물들을 high performance liquid chromatogrphy를 이용하여 분획한 분획물(fraction)들으 $\alpha$-glucosidase(EC 3.2.1.20) 저해제를 탐색하였다. 작약 ethyl acetate 추출물의 11, 12, 18, 19번 분획물들은 20 $\mu\textrm{g}$/mL에서 각각 85%, 84%, 77%, 77%의 강력한 저해 활성을 나타내었으며, 작약 ethyl acetate 추출물의 10~19번 분획을 경구투여한 in vivo당 (maltose) 부하 실험에서도 양성대조군과 비교하여 유의성 있게 22%의 혈당을 낮추었다. 따라서 본 연구의 in vitro와 in vivo 실험을 통하여 확인 된 $\alpha$-glucosidase 저해 활성을 갖는 작약 분획물들은 식후의 혈당상승 작용을 억제하는 새로운 nutraceutical 소재와 항당뇨 신약개발을 위한 탐색자원으로서 매우 가치있는 자원으로 기대된다.

• Journal of Periodontal and Implant Science
• /
• 제28권4호
• /
• pp.611-632
• /
• 1998

Chitosan has been known as a wound healing agent. The purpose of this study was to evaluate the biocompatibility and guided bone regenerative effect of chitosan and chitosan-cellulose membranes. The effects of chitosan and chitosan-cellulose membranes on the growth and survival of human periodontal ligament cells were examined by rapid colorimetric MTT(tetrazolium) assay, and the tissue response and resorption pattern were observed by implanting the membranes into the subcutaneous tissue of the back of rats for 6 weeks. To evaluate the guided bone regenerative potential of membranes, the amount of newly formed bone in the rat calvarial defects(8mm in diameter) was measured by histomorphometry and radiomorphometry 1,2 and 4 weeks after implantation of membranes. Chitosan and chitosan-cellulose membranes showed no adverse effect on the growth and survival of human periodontal ligament cells. When membranes were subcutaneously implanted, inflammatory reaction was observed at 1 week and which gradually subsided 2weeks after implantation. Membranes remained intact throughout the experimental period of 6 weeks. Radiomorphometric analysis of the craniotomy sites revealed that chitosan and chitosan-cellulose membrane implanted sites showed increased radiopacity over control. Statistically significant differences with control were found in chitosan-cellulose membrane implanted group at 2 and 4 weeks, and chitosan membrane implanted group at 4 weeks(P<0.05). Histomorphometric data indicated a pattern of osseous healing similar to radiomorphometric analysis. There was a statistically significant difference between control and chitosan-cellulose membrane implanted group at 4 weeks(P<0.05). These results implicate that chitosan and chitosan-cellulose membrane might be useful for guided bone regeneration.

• Journal of Periodontal and Implant Science
• /
• 제23권1호
• /
• pp.145-158
• /
• 1993

The oral microbiota such as P. gingivalis, P. intermedia and A. actinomycetemcomitans play a primary role in the initiation and progression of the periodontal disease. The purpose of this study was to evaluate the antimicrobial effects and inhibitory effects of honokiol and magnolol on the bacterial collagenase activity, cytotoxicity and cytokine production of periodontopathic microorganisms. The antimicrobial activities of honokiol and magnolol was evaluted with minimum inhibition concentration. Honokiol was more active than magnolol, but less than chlorhexidine on antimicrobial activity. The inhibitory effects of magnolol and honokiol on the collagenolytic activity and cytotoxicity were evaluated using a Collagenokit CLN-100 and rapid colorimetric assay (MTT method) for cellular growth and survival of gingival fibroblast and periodontalligament cell and $[^3H]-thymidine$ incorporation for the gingival epithelial cell. The inhibitory effects on the collagenolytic activity was the highest in chlorhexidine, and the lowest in magnolol. Magnolol had the lowest cytotoxic effect and chlorhexidine had the highest. The inhibitory effects on cytokine production was evaluated using $interleukin-1{\beta}$ ELISA kit (Cistron Biotech.), IL-6, $TNF-{\alpha}$ ELISA kit (Genzyme) and inhibitory effects were higher than bacterial LPS and there is no difference among the honokiol, magnolol and chlorhexidine. From these results, the antimicrobial and antienzymatic activities of honokiol and magnolol were seemed to inhibit bacterial growth and enzyme activities with lesser cytotoxic activities. Therefore, it was suggested that honokiol and magnolol are very effective antimicrobial agents on periodontal pathogens.

• 생약학회지
• /
• 제25권3호
• /
• pp.249-257
• /
• 1994

The cytotoxic and antitumor activity of Perilla frutescens extract on cultured 3T3 fibroblast and skin melanoma cells were evaluated by tetrazolium MTT (MTT) and neutral red (NR) colorimetric assay methods. Lactate dehydrogenase activity was also measured. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and skin melanoma cells. The results were as follows: 1. Water and ether extracts showed a significant cytotoxicity in 3T3 fibroblast and all extracts exhibited a significant anti-tumor activity in skin melanoma cells. Methanol, ethyl acetate and ethanol extracts showed low cytotoxic effects, but exhibited a high anti-tumor activity. 2. The MTT absorbance in 3T3 fibroblast was significantly decreased by treatment with ether, water, chloroform and ethanol extracts and skin melanoma cells was significantly decreased by treatment with all extracts. The difference in MTT absorbance in two cell types was most remarkable when treated with methanol and ethanol extracts. 3. Methanol and ethyl acetate extracts showed the strongest effect in growth inhibition of melanoma cells. These results indicated that methanol extract possessed a low cytotoxicity and a strong anti-tumor activity.

• 한국식품영양학회지
• /
• 제13권5호
• /
• pp.460-464
• /
• 2000

본 연구에서는 전통적으로 담석증, 신장치료 등의 한약제로 많이 사용하는 대왕을 여러 용매를 사용하여 얻은 대왕추출물 분액에 대한 세포독성을 여부를 MTT 정량법, NR 정량법, SRB 정량법을 이용하여 조사하였다. 1. 추출 용매 methylene chloride, ethyl acetate, butanol, water로부터 얻은 대왕추출물 모두 처리농도에 따라 세포에 미치는 영향이 증가하였다. 2. Butanol을 용매로 사용하여 얻은 대왕추출물 분액이 다른 3가지 용매로부터 얻은 대왕추출물보다 세포에 미치는 영향이 크게 나타났고 water를 용매로 사용하여 얻은 추출물이 A498 세포주에 미치는 영향이 가장 낮게 나타났다. 3. Butanol을 추출 용매로 하여 얻은 대왕추출물이 A498 세포주에 미치는 영향이 가장 컸는데 그 추출물에 대한 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.63mg/ml, 0.65mg/ml, 0.68mg/ml이었고, 가장 영향이 적은 water의 경우 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.84mg/ml, 0.82mg/ml, 0.80mg/ml이었다. 4. 정량방법 간의 대왕추출물에 대한 반응은 MTT 정량법이 가장 민감하게 나타났다.

• Nutrition Research and Practice
• /
• 제11권3호
• /
• pp.180-189
• /
• 2017

BACKGROUND/OBJECTIVES: Recent living condition improvements, changes in dietary habits, and reductions in physical activity are contributing to an increase in metabolic syndrome symptoms including diabetes and obesity. Through such societal developments, humankind is continuously exposed to metabolic diseases such as diabetes, and the number of the victims is increasing. This study investigated Cordyceps militaris water extract (CMW)-induced glucose uptake in HepG2 cells and the effect of CMW treatment on glucose metabolism. MATERIALS/METHODS: Colorimetric assay kits were used to determine the glucokinase (GK) and pyruvate dehydrogenase (PDH) activities, glucose uptake, and glycogen content. Either RT-PCR or western blot analysis was performed for quantitation of glucose transporter 2 (GLUT2), hepatocyte nuclear factor 1 alpha ($HNF-1{\alpha}$), phosphatidylinositol 3-kinase (PI3k), protein kinase B (Akt), phosphorylated AMP-activated protein kinase (pAMPK), phosphoenolpyruvate carboxykinase, GK, PDH, and glycogen synthase kinase 3 beta ($GSK-3{\beta}$) expression levels. The ${\alpha}-glucosidase$ inhibitory activities of acarbose and CMW were evaluated by absorbance measurement. RESULTS: CMW induced glucose uptake in HepG2 cells by increasing GLUT2 through $HNF-1{\alpha}$ expression stimulation. Glucose in the cells increased the CMW-induced phosphorylation of AMPK. In turn, glycolysis was stimulated, and glyconeogenesis was inhibited. Furthermore, by studying the mechanism of action of PI3k, Akt, and $GSK-3{\beta}$, and measuring glycogen content, the study confirmed that the glucose was stored in the liver as glycogen. Finally, CMW resulted in a higher level of ${\alpha}-glucosidase$ inhibitory activity than that from acarbose. CONCLUSION: CMW induced the uptake of glucose into HepG2 cells, as well, it induced metabolism of the absorbed glucose. It is concluded that CMW is a candidate or potential use in diabetes prevention and treatment.

• 대한임상검사과학회지
• /
• 제43권2호
• /
• pp.75-81
• /
• 2011

To clarify the oxidative stress of reactive oxygen species (ROS) and the effect of Chrysanthemum morifolium L. (CM) flower extract on the cultured neuroglial cells (C6 glioma) damaged by ROS, cell adhesion effect was measured by colorimetric assay after cultured C6 glioma cells were treated with various concentrations of glucose oxidase (GO) for 5 hours. For the antioxidative effect of CM flower extract, cell adhesion activity (CAA), superoxide dismutase (SOD)-like activity and lactate dehydrogenase (LDH) activity were assessed against GO-induced cytotoxicity on same cultures. In this study, GO remarkably decreased CAA dose-dependently, and the $XTT_{90}$ and $XTT_{50}$ values were measured at 15 mU/mL and 50 mU/mL following the treatment of C6 glioma cells with 5~60 mU/mL of GO. The CM flower extract significantly increased cell adhesion activity damaged by GO-induced cytotoxicity, and it also showed the SOD-like activity and the decrease of LDH activity. From these results, it is suggested that GO was cytotoxic on cultured C6 glioma cells, and CM flower extract showed antioxidative effects as shown by the increased CAA, SOD-like activity and the decrease of LDH activity on GO-induced cytotoxicity on the same cultures.

• Journal of Microbiology and Biotechnology
• /
• 제17권10호
• /
• pp.1607-1615
• /
• 2007

We investigated the applicability of the TEM-l ${\beta}$-lactamase fragment complementation (BFC) system to develop a strategy for the screening of protein-protein interactions in bacteria. A BFC system containing a human Fas-associated death domain (hFADD) and human Fas death domain (hFasDD) was generated. The hFADD-hFasDD interaction was verified by cell survivability in ampicillin-containing medium and the colorimetric change of nitrocefin. It was also confirmed by His pull-down assay using cell lysates obtained in selection steps. A coiled-coil helix coiled-coil domain-containing protein 5 (CHCH5) was identified as an interacting protein of human uracil DNA glycosylase (hUNG) from the bacterial BFC cDNA library strategy. The interaction between hUNG and CHCH5 was further confirmed with immunoprecipitation using a mammalian expression system. CHCH5 enhanced the DNA glycosylase activity of hUNG to remove uracil from DNA duplexes containing a U/G mismatch pair. These results suggest that the bacterial BFC cDNA library strategy can be effectively used to identify interacting protein pairs.