• 생명과학회지
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• 제19권7호
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• pp.871-877
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• 2009

대표적인 histone deacetylase inhibitor 저해제의 일종일 sodium butyrate에 의한 인체백혈병 U937세포의 증식 억제에 관한 기전 연구를 세포주기 조절 측면에서 조사하였다. MTT assay 및 flow cytometry 분석을 통하여 sodium butyrate의 처리 농도 증가에 따른 U937 세포의 증식억제는 세포주기 G1 arrest 및 apoptosis 유발에 의한 것임을 확인하였다. RT-PCR및 Western blotting 결과에서 sodium butrate에 의한 G1 arrest는 세포주기 G1기에서 S기로의 진입에 중요한 역할을 하는 cyclin D1, E, A, cyclin-dependent kinase (Cdk) 4 및 Cdk6발현의 저해와 p21 및 p27과 같은 Cdk inhibitor의 발현 증가와 연관성이 있었다. Sodium butyrate는 또한 retinoblastoma protein (pRB)및 p130 단백질의 인산화를 저해시켰으나, S기 진행에 중요한 전사조절인자인 E2F-1 및 E2F-4의 의 발현에는 큰 영향이 없었다. 그러나 sodium butyrate에 의한 pRB 및 p130단백질의 인산화 저해는 pRB와 E2F-1및 p130과 E2F-4와의 결합력을 증사시켰다. 본 연구의 결과는 U937세포의 증식억제에 pRB/p130 인산화 억제 및 Cdk inhibitors의 발현 증가가 중요한 역할을 하고있음을 보여주는 것으로, sodium butyrate의 항암기전 이해에 중요한 자료가 될 것이다.

• 대한예방한의학회지
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• 제18권1호
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• pp.11-21
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• 2014

Objective : This study set out to combine the treatment efficacy of Taraxacum with Dongchimi fermentation and investigate Taraxacum's effects on protection of liver cell and controlling nitric oxide(NO) through experiments, thus checking whether it had values as a physiological active matter. The experimental materials include Taraxacum Dongchimi (TD) and Taraxacum fermented by Dongchimi (TDF). As for methodology, experiments were carried out to compare TD and TDF in components, protection effects for liver cells, anticancer effects on liver cells, and protection effects for brain cells in the aspects of liver function and immunity enhancement. Method : The experimental materials include Taraxacum Dongchimi (TD) and Taraxacum fermented by Dongchimi (TDF). As for methodology, experiments were carried out to compare TD and TDF in components, protection effects for liver cells, anti-cancer effects on liver cells, and protection effects for brain cells in the aspects of liver function and immunity enhancement. Results : As shown in the chromatogram results, each valid component content increased in Taraxacum fermented by Dongchimi (TDF) for each time section. Of them, the valid component content at 36.80 minutes was approximately 2.7 times higher in TDF at 21.8% than in Taraxacum Dongchimi (TD) at 8.28%. TDF generated more excellent protection effects against the toxicity that caused oxidative damage to the liver cell(HepG2) with t-BHP than TD. The survival rate was low in TD of $100{\mu}g/m{\ell}$ and $300{\mu}g/m{\ell}$ and increased to 23.3% in TDF of $100{\mu}g/m{\ell}$. The survival rate was the highest at $300{\mu}M$ with a significant difference of 68.1%(P<0.05). Both TD and TDF showed effects of controlling nitric oxide production according to concentration with TDF recording a higher rate of controlling nitric oxide production than TD. There were significant differences(P<0.05) in the effects of controlling nitric oxide production at 200 ug/ml, 400 ug/ml in both groups. Especially the result TDF of $400{\mu}g/m{\ell}$ was thus similar to those of butein, the positive control group. Conclusion : The result of this studies is that Taraxacum fermented by Dongchimi (TDF) increased the valid component content compared with the simple mixture(TD). The findings clearly show that it is a material with the effects of improving immunity and liver cell protection. If fermentation methods are further developed to use it as a functional material, it will be subject to more opportunities of being used in other functional foods and make a contribution to integrated medicinal food development.

• 동의생리병리학회지
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• 제21권4호
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• pp.856-859
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• 2007

We have investigated the antimetastatic and antitumor effects of Ginsenoside Rh2 and ${\beta}-glucan$ unsing an experimental metastatic mouse model intravenously injected with B 16 melanoma F 10 cells. Animal groups are divided into six groups according to the dosage of drug administration and the kind of drugs. The groups are control, ${\beta}-glucan$ with 50, 100 and 200 mg/kg, Geinsenoside Rh2 50 mg/kg, and ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg. Oral administration of various concentration of ${\beta}-glucan$( 50, 100, and 200 mg/kg) were reduced the lung- metastatics induced by metastatic B16 melanoma F 10 cells injection with a dose dependent manner in the syngenic mice. At same dosage group, Ginsenoside Rh2 (50 mg/kg) has more antimetastatic effect than the ${\beta}-glucan$(50 mg/kg). The highest antimetastatic effects was observed in the ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group and has a similar tendency in the anti-tumor effects, including decrease of the average tumor weight and increase of the average survival rate. There are no differences of the average tumor weights were apparent in the ${\beta}-glucan$ groups, however there were little decrease of the average tumor weight in Ginsenoside 50 mg/kg group and ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group than that of the control group. The rate of average survival rate in the ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group, ${\beta}-glucan$ 200 mg/kg, ${\beta}-glucan$ 100 mg/kg and ${\beta}-glucan$ 50 mg/kg, and Ginsenoside 50 mg/kg groups were highly in order. These data suggest that antimetastatic and antitumor effect of combination of Ginsenodide Rh2 and ${\beta}-glucan$ be the highest in this study.

• 한국식품위생안전성학회지
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• 제27권4호
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• pp.406-414
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• 2012

본 연구에서는 fucoidan이 macrophage의 활성과 항암효과를 확인하기 위하여 수행되었다. Fucoidan을 Raw 264.7 세포에 처리한 후 MTT assay로 측정한 결과 고농도 $100{\mu}g/mL$까지 세포독성은 없었으며, NO 및 TNF-${\alpha}$의 분비를 농도 유의적으로 증가시켰다. 또한 AGS 위암 세포 성장저해효과를 확인하기 위하여 MTT assay를 하였고 그 결과 농도와 시간 의존적으로 암 세포 성장이 유의적으로 감소하였다. Apoptosis로 인해 암 세포 성장이 감소하였는지 확인하기 위하여 DAPI 염색을 한 결과 apoptotic body와 세포질 응축이 시간 의존적으로 증가하는 것을 확인하였다. 또한 fucoidan은 미토콘드리아의 투과율을 향상시키며 미토콘드리아에서 방출되는 cytochrome c의 발현을 증가시켰다. Western blotting의 결과 시간 의존적으로 anti-apoptotic 분자인 Bcl-2와 XIAP 발현 감소와 반대로 pro-apoptotic 분자인 Bax 발현이 증가하였다. Cleaved-caspase-9의 발현이 증가하였으며 Akt의 인산화는 시간 의존적으로 감소하였다. Caspase 억제제인 z-VAD-FMK 처리 시 Bax, caspase-9의 발현을 감소시켜 apoptosis 유도를 억제하였으며 이러한 결과는 caspase가 apoptosis 유도에 중요한 역할을 하는 것으로 나타낸다. 본 실험의 AGS 위암 세포주에서 대조군에 비하여 fucoidan 처리군에서 면역세포 활성 및 AGS 위암 세포주에서 caspase 활성을 통해 apoptosis를 유도하는 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.531-540
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• 2022

Due to the high incidence of malignant melanoma, the establishment of in vitro models that recapitulate the tumor microenvironment is of great biological and clinical importance for tumor treatment and drug research. In this study, 3D printing technology was used to prepare GelMA/PEGDA composite scaffolds that mimic the microenvironment of human malignant melanoma cell (A375) growth and construct in vitro melanoma micro-models. The GelMA/PEGDA hybrid scaffold was tested by the mechanical property, cell live/dead assay, cell proliferation assay, cytoskeleton staining and drug loading assay. The growth of tumor cells in two- and three-dimensional culture systems and the anti-cancer effect of luteolin were evaluated using the live/dead staining method and the Cell Counting Kit-8 (CCK-8) method. The results showed a high aggregation of tumor cells on the 3D scaffold, which was suitable for long-term culture. Cytoskeleton staining and immunofluorescent protein staining were used to evaluate the degree of differentiation of tumor cells under 2D and 3D culture systems. The results indicated that 3D bioprinted scaffolds were more suitable for tumor cell expansion and differentiation, and the tumor cells were more aggressive. In addition, luteolin was time- and dose-dependent on tumor cells, and tumor cells in the 3D culture system were more resistant to the drug.

• 대한본초학회지
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• 제32권5호
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• pp.27-38
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• 2017

Objective : In the present study, we examined whether Canavalia gladiata D.C. (CG) and Arctium lappa L., Redix (AL) mixture (CGAL), their components, lupeol and chicoric acid, regulate immune system and suppress the tumor in vitro and in vivo. Methods : LPS-induced reactive oxygen species (ROS) and nitric oxide (NO) were measured after treatment with CG extract (CGE), CGAL, lupeol, chicoric acid and lupeol and chicoric acid mixture (lupeol+CA) in Raw264.7 cell. To determine the effect of CGE on immune responses, immune cell population and IgG production were assessed in mice. To investigate the effect of CGAL and their component on anti-tumor activity, tumor volume and weight were measured, cell cycles and immune cell population were analyzed in MC38 injected tumor bearing mice. Also, NK cell activity was determined in splenocyte isolated from tumor bearing mice. Results : CGE, CGAL, lupeol, chicoric acid and lupeol+CA decreased the LPS-induced ROS and NO production without cell toxicity in RAW264.7 cells. CGE increased the immune cell populations of $CD4^+T$, $CD8^+T$ and macrophages in various immune organ of mice. In tumor bearing mice, CGAL, lupeol, chicoric acid and lupeol+CA suppressed tumor volume and weight. In cell cycle analysis, they decreased the percentages of S phase. In addition, CGAL, lupeol, chicoric acid and lupeol+CA immune cell populations of $CD4^+T$, $CD8^+Tcell$, NK cell and macrophage in tumor as well as NK cell activity. Conclusion : CGAL and its compounds may enhance immune responses and suppress tumor growth, and may be capable of developing health functional foods.

• 생명과학회지
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• 제31권5호
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• pp.475-480
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• 2021

국내에서 2016년 흰점박이꽃무지 등 일부 곤충이 식품원료로 등록되면서 곤충사육 농가가 폭발적으로 증가하고 있다. 하지만 곤충사육 농가가 보유하고 있는 사육기술, 병해충 관리기술이 농가별로 차이가 있고, 대량으로 곤충을 사육하면서 높은 사육밀도, 병충해 감염 등으로 인해 곤충이 대량 폐사하게 됨으로써 곤충사육 농가에 심각한 경제적 손실을 입고 있다. 이에 본 연구에서는 다양한 영양성분을 함유할 뿐만 아니라 항균, 항염증 등의 효과를 가진 새싹보리를 흰점박이꽃무지 유충의 주 먹이원인 발효톱밥에 첨가하여 주 먹이원 첨가제로써 적용 가능성 및 곤충병원성 세균에 대한 항균 효과를 확인하고자 하였다. 흰점박이꽃무지 유충의 주 먹이원에 5%와 10% 새싹보리를 첨가했을 때 유충의 증체율이 19.2%와 23.1%로 높았고, 사육 기간이 대조군 대비 단축된 것으로 조사되었다. 또한 새싹보리는 곤충병원성 세균 Serratia marcescens의 성장을 저해하고, 세균에 감염된 흰점박이꽃무지 유충의 생존율과 증체율이 건강한 유충의 수준으로 회복되는 경향을 보였다. 뿐만 아니라 새싹보리를 첨가한 먹이원으로 사육한 유충의 영양성분 분석 결과, 조단백질, 조지방, 구리, 아연, 칼륨 등 미네랄 및 아미노산 함량이 대조군보다 높게 나타났다. 결론적으로 흰점박이꽃무지 유충의 주 먹이원에 새싹보리를 첨가한다면 단기간에 대량생산이 가능하고, 곤충병원성 세균 감염에 의한 폐사를 효율적으로 예방할 수 있어 안정적인 곤충 생산이 가능할 것으로 판단된다.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.33-45
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• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

• 한국식품영양과학회지
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• 제38권1호
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• pp.25-31
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• 2009

본 연구는 잔대의 돌연변이원성, 항돌연변이원성, 세포독성, 항종양 효과를 조사하기 위해서 수행되었다. 잔대를 70% 에탄올로 추출하여 추출용매에 따라 핵산, 클로로포름, 에틸아세테이트, 부탄올과 물층으로 분획하였다. Ames test, SRB assay와 종양 억제실험 방법을 사용하여 실험하였다. Ames test결과, 잔대 에탄올 추출물과 그 분획물들은 돌연변이원성을 나타내지 않았을 뿐만 아니라, MNNG와 4NQO로 돌연변이를 일으켰을 때 높은 항돌연변이 효과를 나타내었다. 잔대 에틸아세테이트 분획물은 S. Typhimurium TA98를 4NQO로 돌연변이를 유도하였을 때 66.5%의 억제율을 나타내었으며, S. Typhimurium TA100에서 MNNG와 4NQO로 돌연변이를 유도했을 때는 83.3%와 75.1%의 억제율을 나타내었다. 잔대 추출물 및 그 분획물들의 암세포 성장 억제효과를 살펴보기 위해 인간 자궁암세포 (HeLa), 인간 간암세포(Hep3B), 인간 유방암세포(MCF-7), 인간 위암세포(AGS), 인간 폐암세포(A549)와 인간 신장정상세포(293)를 사용하였다. 잔대 에틸아세테이트 분획물을 1 mg/mL를 처리하였을 때 각각 79.9%(HeLa), 74.9% (Hep3B), 66.0%(MCF-7), 71.0%(AGS)와 74.3%(A549)의 가장 높은 억제활성을 나타내었다. 반면에 인간 정상 신장세포에서는 $3{\sim}36%$의 세포독성을 나타내었다. In vivo에서 잔대 추출물의 항암효과를 시험하기 위하여 Balb/c 마우스에 sarcoma-180 종양세포로 고형암을 유발시켰다. 그 결과 잔대 에틸아세테이트 층의 최고농도 50 mg/kg에서 37.2%의 고형암 성장 억제효과를 나타내었고, 이는 모든 처리군 중가장 높은 억제율이었다.

• 생명과학회지
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• 제29권3호
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• pp.337-344
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• 2019

본 연구에서는 택란 잎에 대한 다양한 생리활성을 규명하기 위하여 택란 잎을 극성이 다른 용매로 분획하여 분획물의 플라보노이드 함량을 측정하고 택란 잎 분획물에 의한 라디칼 소거활성과 세포독성 효과를 규명하고자 하였다. 택란 잎 85% MeOH 및 n-BuOH분획물들의 총 플라보노이드 함량은 각각 $41.5{\pm}0.00mg/g$ 및 $77.2{\pm}0.01mg/g$으로 n-BuOH 분획물의 총 플라보노이드 함량이 가장 높았음을 알 수 있었다. DPPH assay에서 택란 잎 n-BuOH 분획물의 활성산소 소거능이 가장 우수하였고 다음으로 85% MeOH 분획물에 의한 소거활성이 높았다(p<0.05). 각 분획물들의 $IC_{50}$값은 n-Hexane 9.76 mg/ml, 85% MeOH 0.50 mg/ml, n-BuOH 0.03 mg/ml 및 water 0.78 mg/ml로 나타났다. 또한 택란 잎 85% aq. MeOH 및 n-BuOH 분획물들에 의한 ABTs+소거활성은 각각 89.3% 및 92.1%이었으며 대조군인BHT와 L-ascorbic acid와 유사한 값을 나타내었다(p<0.05). ABTs+ assay에서 각 분획물들의 $IC_{50}$값은 n-Hexane 0.96 mg/ml, 85% MeOH 0.16 mg/ml, n-BuOH 0.01 mg/ml 및 water 0.31 mg/ml로 나타났다. 택란 잎 n-Hexane, 85% MeOH, n-BuOH 및 water 분획물들은 농도의존적으로 암세포들에 대한 세포독성 효과를 나타내었으며 분획물들 중 특히 n-Hexane과 85% MeOH 분획물들에 의한 세포독성 효과가 가장 높았다. AGS 암세포에 대한 택란 잎 n-Hexane, 85% MeOH, n-BuOH 및 water 분획물들의 $IC_{50}$ 값은 각각 0.18, 0.03, 0.62 및 0.12 mg/ml이었고 HT-29 암세포에 대한 $IC_{50}$ 값은 각각 0.26, 0.16, 0.60 및 1.14 mg/ml이었다. HT-1080 암세포에서 택란 잎n-Hexane, 85% MeOH, n-BuOH 및 water 분획물들의 $IC_{50}$ 값은 각각 0.34, 0.14, 0.48, 및 1.05 mg/ml이었다. 따라서 본 연구결과로부터 택란 잎 n-BuOH 분획물은 자유라디칼 소거활성에 효과적이었고 n-Hexane과 85% MeOH 분획물들은 높은 세포독성 효과를 나타내었으며 향후 택란 잎을 이용한 식품개발을 위한 기초자료를 제시하고자 한다.