• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7849-7855
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• 2014

Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

• 대한임상검사과학회지
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• 제49권2호
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• pp.150-160
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• 2017

아토피피부염은 만성 염증성 피부질환으로 습진성병변, 피부건조 및 가려움증의 특징적 증상을 보이는 질환이다. 현재 사용되고 있는 아토피피부염 치료약제들은 오래 사용시 여러 부작용이 있어서 이를 대신할 새로운 대체 치료법의 개발이 요구되고 있다. Curcumin은 항염, 항알러지와 같은 효과를 나타내는 천연 폴리페놀(polyphenol)이며, 발광다이오드(light emitting diode, LED) 광치료는 아토피피부염과 상처치유에 효과가 있는 것으로 알려져 있다. 본 연구는 DNCB로 유도한BALB/c 마우스의 아토피피부염에서 curcumin 투여와 630 nm LED 조사를 병용하여 그 치료 효과를 높일 수 있는가를 조사하기 위해 시도하였다. 아토피피부염의 치료 효과를 조사하기 위해 modified SCORAD 지수, 조직병리학적 분석, 면역조직화학염색 및 TUNEL 검사를 사용하였다. 그 결과 curcumin 투여와 630 nm LED 조사가 각각 SCORAD 지수, 비만세포 및 탈과립세포의 수, 호산구 수, 표피세포의 증식능 및 자멸세포의 수를 의미 있게 감소시키고, 비만세포의 탈과립을 억제하여 섬유모세포에 의한 콜라겐 생성을 의미 있게 감소시켰으며, curcumin 투여와 LED 조사를 병용하였을 경우 단일 처리군에 비해 이들 수치가 의미 있게 감소하는 것을 확인하였다. 이러한 결과는 아토피피부염 치료에 curcumin 투여와 LED 조사가 각각 효과가 있음을 의미하며, 병용 치료 시 그 효과가 좀 더 향상된다는 것을 의미하는 것으로, 아토피피부염의 대체치료 전략으로 curcumin 투여와 630 nm 조사의 병용 치료를 제안한다.

• Asian-Australasian Journal of Animal Sciences
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• 제24권4호
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• pp.485-492
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• 2011

The influence of high temperature on mammary cell turnover during the dry period is still unclear. The objective of this study was to investigate mammary cell turnover and p53 protein expression in the mammary tissue under high temperature conditions. Mammary gland biopsy samples from 8 dairy cows were obtained at 7, 25, 40, and 53 d during the dry period in summer or spring (n = 4, each season). Cell cycle, cell turnover, and p53 protein expression were analyzed by flow cytometry. During the dry period in summer, the percentage of mammary epithelial cells in the G0/G1 phase was the highest, but those in the S and G2/M phases were lower. However, the proportion of cells in the different stages of the cell cycle was not significantly different among the different biopsy time points, except in the G2/M phase. Under different temperature conditions, the cells were significantly different in their apoptotic rate and proliferation index; moreover, the tendencies of these indicators to change significantly differed. In general, the samples under high temperature conditions showed significantly lower apoptotic rates and proliferation indices. Under high temperature conditions, the apoptotic rate and proliferation index were the lowest (2.17% and 3.26%, respectively) at day 40, and the highest at day 53 (3.67% and 4.61%, respectively). However, under normal temperature conditions, the values of these indicators were the lowest (7.60% and 5.54%, respectively) at day 7, and almost the highest at day 25 (12.85% and 6.47%, respectively). Moreover, p53 protein expression was significantly higher under high temperature conditions than under normal temperature conditions, except at day 25. The level of p53 protein was the lowest (13.10%) under high temperature conditions at day 25, but was the highest (26.07%) under normal temperature conditions. Our findings suggest that high temperature delayed the G2/M phase of the cell cycle and the cell turnover rate, but remarkably increased p53 protein expression. Thus, the results indicate that high temperature extends the recovery period of mammary epithelial cells.

• 대한방사선기술학회지:방사선기술과학
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• 제39권3호
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• pp.451-459
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• 2016

방사선 사고 및 방사선 치료 등 방사선에 의한 과피폭의 피해를 줄이기 위해 방사선방호제 개발연구는 활발히 진행되고 있다. 이에 화학적 합성물이 아닌 항산화, 항암, 면역 증강에 효과적인 것으로 알려진 백하수오를 이용하여 방사선방호효과를 확인하였다. 백하수오 에탄올추출물을 Sprague Dawley Rat (SD Rat)에 14일간 1일 1회 경구 투여하고, 7 Gy X-ray를 조사한 후 1일, 4일, 7일, 21일의 시간 변화에 따른 혈구성분, 비장 지수의 변화 및 간과 자궁의 조직변화를 관찰하였다. 실험결과 백하수오 에탄올추출물을 섭취한 실험군의 백혈구 수치(p < 0.05)와 비장 지수(p < 0.05)가 대조군보다 회복이 빠른 것을 확인하였다. 간 조직에서는 핵의 응축, 세포질의 팽창, 염증세포의 침윤이 감소하였으며, 자궁샘 조직은 세포고사가 감소한 것을 확인하였다. 위의 결과를 토대로 백하수오 에탄올추출물은 방사선 조사에 따른 혈구 및 장기의 피해를 줄일 수 있는 새로운 방사선방호제로써 유용할 것으로 기대되며, 방사선 사고와 같은 비상관리 분야에 적절한 시사점을 제공할 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제50권1호
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• pp.44-52
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• 2023

Objective: The DNA integrity of spermatozoa that attach to fallopian tube (FT) cells is higher than spermatozoa that do not attach. FT epithelial cells can distinguish normal and abnormal sperm chromatin. This study investigated the effects of sperm with a high-DNA fragmentation index (DFI) from men with unexplained repeated implantation failure (RIF) on the Toll-like receptor (TLR) signaling pathway in human FT cells in vitro. Methods: Ten men with a RIF history and high-DFI and 10 healthy donors with low-DFI comprised the high-DFI (>30%) and control (<30%) groups, respectively. After fresh semen preparation, sperm were co-cultured with a human FT epithelial cell line (OE-E6/E7) for 24 hours. RNA was extracted from the cell line and the human innate and adaptive immune responses were tested using an RT2 profiler polymerase chain reaction (PCR) array. Results: The PCR array data showed significantly higher TLR-1, TLR-2, TLR-3, TLR-6, interleukin 1α (IL-1α), IL-1β, IL-6, IL-12, interferon α (IFN-α), IFN-β, tumor necrosis factor α (TNF-α), CXCL8, GM-CSF, G-CSF, CD14, ELK1, IRAK1, IRAK2, IRAK4, IRF1, IRF3, LY96, MAP2K3, MAP2K4, MAP3K7, MAP4K4, MAPK8, MAPK8IP3, MYD88, NFKB1, NFKB2, REL, TIRAP, and TRAF6 expression in the high-DFI group than in the control group. These factors are all involved in the TLR-MyD88 signaling pathway. Conclusion: The MyD88-dependent pathway through TLR-1, TLR-2, and TLR-6 activation may be one of the main inflammatory pathways activated by high-DFI sperm from men with RIF. Following activation of this pathway, epithelial cells produce inflammatory cytokines, resulting in neutrophil infiltration, activation, phagocytosis, neutrophil extracellular trap formation, and apoptosis.

• 한국식품영양과학회지
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• 제41권8호
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• pp.1079-1085
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• 2012

본 연구는 녹두묵을 이용한 항비만 임상선행연구를 바탕으로 녹두에 풍부한 vitexin의 항비만 효과의 기작을 살펴보고자 하였다. 녹두 vitexin은 $200{\mu}M$ 농도까지 3T3-L1 지방 전구세포의 독성효과는 없었으며 50, 100, $200{\mu}M$ 농도에서 지방합성이 억제되었고 $100{\mu}M$ 농도에서 지방분해효과로 추측할 수 있는 결과가 나타났다. 또한 $200{\mu}M$에서 염증성 아디포사이토카인 유전자발현을 증폭시키는 상위신호체계(p38, ERK)가 억제된 반면 AMPK의 경우는 $100{\mu}M$ 농도까지 증가하였고 $PPAR{\gamma}$ 단백질 발현도 $200{\mu}M$에서 증폭하였으며 TNF-${\alpha}$와 aP2 mRNA 발현은 $25{\mu}M$에 증가하였다가 감소하였다. 이러한 결과를 바탕으로 결론을 내리면 vitexin $100{\sim}200{\mu}M$ 사이의 농도가 항비만 기작을 규명하는 in vivo 실험을 위한 적절한 농도로 여겨진다. 그러나 본 연구에서는 vitexin 처리에 따른 C/EBP, SREBP1, $PPAR{\gamma}$ mRNA 농도의 변화는 관찰되지 않아 3T3-L1 세포자살(apoptosis) 신호체계 및 mTOR와의 연계성체계 혹은 기타 adipogenesis 억제 유전자로 SIRT1 등의 연구가 향후에 더 필요할 것으로 사료된다.

• 대한소아치과학회지
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• 제29권2호
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• pp.243-254
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• 2002

소아치과 임상에 흔히 사용되는 3종의 금속재에 의한 인체 섬유모세포(HGF-1)를 이용하여 세포독성에 대한 세포적 및 분자적 접근을 시도하였다. 세포의 성장과 증식 및 세포사, 그리고 이들과 관련된 세포내 신호전달물질 발현의 변화를 보고자 하였고, 세포사는 screening한 후 세포사의 유형(necrosis/apoptosis)을 규명하고자 하였다. HGF-1은 DMEM으로 계대배양한 후 유지하였으며, 사용한 금속재는 stainless steel crown (R), 교정용 band(B), 교정용 wire(W)의 세 종류였다. HGF-1세포의 증식에 대하여 기본적으로 cell count를 하였고, 생존세포 count를 위하여 대조군에 대한 실험군의 상대증식도를 계산하였다. 세포수는 배양일이 지남에 따라 증가하여 7일째에 최고를 나타내었고, 11일째에는 감소하였으며, 대조군과 실험군 사이의 통계적 유의한 차이는 관찰되지 않았다(p>0.05). 대조군에 대한 실험군의 상대증식도 또한 유의한 차이는 없었다(p>0.05). 세포사에 대한 형태적 소견은 괴사(necrosis)를 나타내었다. PCNA lableing index에 대한 대조군과 실험군간에 유의한 차이는 없었다(p>0.05). 신호전달 관련 MAP kinase인 ERK1 및 ERK2, p38, JNK는 배양일의 경과에 따른 발현의 변화는 관찰되었으나, 대조군과 각 실험군 간에 차이는 관찰되지 않았다. 결론적으로 본 연구대상인 stainless steel crown, 교정용 band 와 wire, 세 종류의 소아 치과용 금속재는 단기간의 인체섬유모세포를 이용한 세포친화성 검정에 있어서 세포적, 분자적 위해 양상을 나타내지 않았다.

• 한국수정란이식학회지
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• 제22권2호
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• pp.81-87
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• 2007

본 연구에서는 수컷 랫드에 DEHP를 투여하여 실험적으로 생식 독성을 유발하고, vitamin E와 catechin을 단독 및 병용 투여하여 수컷 랫드에서 정소의 조직학적 변화, 정액 특성의 변화 및 정자의 운동성 변화 등을 조사하여 그 예방 효과를 알아보고자 하였다. DEHP를 투여한 군은 대조군에 비해 정자농도, 정자 생존율, 정상 정자율이 감소하였다. DEHP에 의한 생식 독성을 예방하기 위해 vitamin E와 catechin을 단독 또는 병행 투여한 결과, DEHP를 투여한 군에 비하여 정자 농도, 정자 생존율, 정상 정자율이 증가하였다. 이러한 결과를 종합해 볼 때, catechin과 vitamin E의 투여는 DEHP에 의한 생식 독성을 예방하는 효과가 있는 것으로 사료된다.

• 대한한의학방제학회지
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• 제25권4호
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• pp.509-526
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• 2017

Background and objectives : Soeumin Bojungykgi-tang (seBYTE) has been used to supplement qi in Korean medicine. It has been demonstrated to possess various biological functions such as anti-cancer, anti-aging and anti-inflammatory effects. The present study evaluated the protective roles of seBYTE in hepatotoxic in vitro and in vivo model. Methods : To investigate cytoprotective effect of seBYTE, HepG2 cells were pretreated with seBYTE and then subsequently exposed to $10{\mu}m$ AA for 12 h, followed by $5{\mu}m$ iron. Cell viability was examined by MTT assay, and expression of apoptosis-related proteins was evaluated by immunoblot analysis. For responsible molecular mechanisms, ROS production, GSH contents, and mitochondrial membrane potential were measured. In addition, hepatoprotective effect of seBYTE in vivo was assessed in $CCl_4$-induced animal model. Results : seBYTE prevented AA + iron-induced cytotoxicity in concentration dependent manner. In addition, ROS production, GSH depletion, and mitochondrial dysfunction induced by AA + iron were significantly reduced by seBYTE pretreatment. Furthermore, seBYTE recovered expression of the pro-apoptotic proteins such as PARP and pro-caspase-3. In animal experiment, plasma ALT and AST levels were significantly elevated in $CCl_4$ treatment, but seBYTE significantly decreased the ALT and AST levels. Moreover, seBYTE alleviated the numbers of histological activity index, percentages of degenerative regions, degenerated hepatocytes, infiltrated inflammatory cells, nitrotyrosine- and 4-hydroxynonenal-positive cells in liver. Conclusions : These results showed that hepatoprotective effect of seBYTE against on $CCl_4$-induced hepatic damages is partly due to antioxidative and anti-apoptotic process.

• Toxicological Research
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• 제28권1호
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• pp.39-49
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• 2012

We investigated the effect of zinc on the formation of colonic aberrant crypt foci induced by azoxymethane (AOM) followed by dextran sodium sulfate (DSS) in mice with high iron diet (HFe; 450 ppm iron). Sixweek old ICR mice were fed on high iron diets with combination of three different levels of zinc in diets, low-zinc (LZn; 0.01 ppm), medium-zinc (MZn; 0.1 ppm), and high-zinc (HZn; 1 ppm) for 12 weeks. Animals were received weekly intraperitoneal injections of AOM (10 mg/kg B.W. in saline) for 3 weeks followed by 2% DSS (molecular weight 36,000~50,000) in the drinking water for a week. To confirm the iron storage in the body, the hepatic iron concentration has been determine chemically and compared with histological assessment visualized by Prussian blue reaction. Aberrant crypt (AC) and aberrant crypt foci (ACF) were analyzed in the colonic mucosa of mouse fed high dietary iron. Superoxide dismutase (SOD) activity and thiobarbituric acid-reactive substances (TBARS) level were also investigated. Apoptosis in the preneoplastic lesion was determined by terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL). In addition, immunohistochemistry of ${\beta}$-catenin was also performed on the mucous membrane of colon. The number of large ACF (${\geq}4$ AC/ACF), which possess greater tumorigenic potential, was significantly lower in MZn and HZn groups compared with LZn group. Cytosolic SOD activity in the liver was significantly higher in HZn group compared with LZn group. Hepatic MDA level was decreased significantly in HZn group compared with MZn and LZn groups. Apoptotic index was significantly higher in HZn group. Taken together, these findings indicate that dietary zinc might exert a protective effect against colonic preneoplastic lesion induced by AOM/DSS in ICR mice with high iron status, and suggest that dietary supplement of zinc might play a role in suppressing colon carcinogenesis in mice.