• 제목/요약/키워드: ITS-PCR

검색결과 2,037건 처리시간 0.034초

Naringenin이 NF-$\kappa$B, AP-1 억제를 통한 MMP-9 활성 및 발현 억제 효과 (Inhibitory Effect of Naringenin on MMP-9 Activity and Expression in HT-1080 Cells)

  • 채수철;고은경;서은선;유근창;나명석;김인숙;이종빈
    • 환경생물
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    • 제27권1호
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    • pp.58-65
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    • 2009
  • Naringenin은 flavonoid구조의 감귤류 과피에 다량 함유되어 있으며 항암 및 항산화 등의 다양한 생리활성을 가지는 것으로 보고되었다. 이에 본 연구에서는 HT-1080 섬유육종세포의 전이에 대한 영향을 조사하였다. 먼저 Naringenin이 암세포의 이동에 미치는 영향을 알아보기 위해 migrationassay를 한 결과, Naringenin이 암세포의 이동을 농도 의존적으로 억제시켰다. 암의 전이에 있어서 매우 중요한 역할을 하는 단백질분해효소인 matrix metalloproteinase-9 (MMP-9)의 활성도를 측정하기 위해 gelatin zymography를 한 결과, Naringenin이 PMA에 의해 증가된 MMP-9의 효소 활성도를 농도 의존적으로 감소시켰다. 또한 MMP-9와 TIMP-1의 유전자 발현에 대한 Naringenin의 영향을 RT-PCR방법으로 조사한 결과, Naringenin이 PMA에 의해 증가된 MMP-9의 mRNA발현을 농도 의존적으로 감소시켰으나 TIMP-1의 mRNA발현에는 변화가 없었다. MMP-9발현 감소에 관여하는 전사조절인자를 확인하기 위해 promoterassay를 한 결과, Naringenin이 PMA에 의해 증가된 MMP-9 및 NF-$\kappa$B, AP-1의 Promoter활성을 농도 의존적으로 감소시 켰다. 또한 MMP-9의 전자조절인자인 NF-$\kappa$B와 AP-1의 결합 활성도를 electrophoretic mobility shift assay로 확인한 결과 Naringenin이 PMA에 의해 증가된 NF-$\kappa$B와 AP-1의 결합 활성도를 농도 의존적으로 감소시켰다. 결론적으로 Naringenin이 전사조절인자인 NF-$\kappa$B와 AP-1의 활성을 억제함으로써 MMP-9의 유전자 발현 및 효소 활성을 억제하고 그 결과 암세포의 이동과 침윤을 억제하는 것을 알 수 있다.

연교(連翹)가 만성 비세균성 전립선영 Rat의 염증발현인자 및 세포조직 변화에 미치는 영향 (The Effects of Forsythiae Frucus on Inflammatory Genes and Cyto-pathological Alterations in Chronic Non-Bacterial Prostatitis Rat Model)

  • 이진신;안영민;안세영;두호경;이병철
    • 대한한방내과학회지
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    • 제27권3호
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    • pp.639-652
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    • 2006
  • Objective : The etiology of chronic prostatitis is likely multifactorial, resulting from either a cascade of events after an initiating factor or from a variety of etiologic mechanisms. There is substantiating evidence to support the role of the inflammatory responses in its pathogenesis, and the clinical value in the evaluation of therapeutic efficacy. Forsythiae Frucus has been traditionally used in treatment of inflammatory diseases, including of prostatitis and urinary tract inflammation. In this study, we investigated the effects of Forsythiae Frucus on inflammatory cytokines and cyto-pathological alternation in the rat model of chronic non-bacterial prostatitis induced by castration and $17{\beta}$-estradiol treatment. Methods : Two-month-old rats were treated with $17{\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis. which is similar to human chronic prostatitis in histopathological profiles. Forsythiae Frucus as an experimental specimen, and testosterone as a positive control, were administered orally. The prostates were evaluated by histopathologlcal parameters including the epithelial score and epithelio-stromal ratio for glandular damage. and the expression of inflammatory cytokine genes including interleukin (IL)-$1{\beta}$, IL-5, IL-12, tumor necrosis factor (TNF)-$\alpha$. eotaxin, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(cox-2). Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation. the rats treated with Forsythiae Frucus showed a diminished range of tissue damage. Epithelial score was improved in the Forsythiae Frucus group over that of the control (P<0.05). The epithelia-stromal ratio was lower in the Forsythiae Frucus group when compared to that of the control (P<0.05). In the reverse transcription-polymerase chain reaction (RT-PCR) of inflammatory cytosine genes. Forsythiae Frucus inhibited the expression of IL-$1{\beta}$, TNF-$\alpha$, iNOS, cox-2 genes, while it modulated the expression of IL-5, which is an anti-inflammatory cytokine. Conclusions : These findings suggest that Forsythiae Frucus may protect the glandular epithelial cells and also inhibit stromal proliferation in association with the immune modulation including the suppression of inflammatory cytokines and increase of anti-inflammatory cytokines. From theses results. we suggest that Forsythiae Frucus could be a useful remedy agents for treating chronic non-bacterial prostatitis.

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사매가 위암세포에 미치는 영향 (The Effects of Duchesnea india(Audra.) Foche. on Human Stomach Cancer Cells)

  • 류봉하;김진성;윤상협;류기원;홍상선
    • 대한한방내과학회지
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    • 제24권2호
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    • pp.290-299
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    • 2003
  • Background : Nowadays many researches about it s cure are going on world widely since cancer is one of the most human health threatening diseases. In Chinese and North Korean medicine, Duchesnea india(Audra.) Foche. is practically used to treat many kinds of cancer, but in Korea it is rarely used. So, we need to scientifically identify anti-tumor effects of Duchesnea india(Audra.) Foche. Objective : We are aimed to identify anti-tumor effects of Duchesnea india(Audra.) Foche. on the stomach cancer cells through molecular biologic methods. Material & Methods : We used AGS as stomach cancer cells from American Type Culture Collection. We added the boiled extract of Duchesnea india(Audra.) Foche. $5{\mu}l$(Sample I), $10{\mu}l$(Sample II) to cultural media(ml)for 0,6, 12, 24, 48 hours. We measured the killing effect on stomach cancer cells through Tryphan blue exclusion test and the suppressive effect on viability of stomach cancer cells via MTT assay. the quantitative RT-PCR was used to examine their effect on the revelation of Bcl-2, Bcl-XL, and Bax, which are genes related to apoptosis. We measured change of mitochondria membrane permeability and membrane potential via flow cytometry. Result : 1. The killing effect on stomach cancer cells showed that each test groups killed more stomach cancer cells than the control group with a time(6 hours later) and density dependent manner, which was statistical significance. 2. The suppressive effect on viability of stomach cancer cells showed that each test groups had more suppressive effects on viability of stomach cancer cells than the control group with a time(6 hours later), which was statistical significance. 3. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 and Bcl-XL decreased with a density manner which was statistical significance. but the revelation of Bax was not changed with statistical significance. 4. As a result of this test, Duchesnea india(Audra.) Foche. caused apoptosis by decreasing the absorbance of mitochondria with statistical significance. and also induced apoptosis by decreasing the membrane potential of mitochondria. Conclusion : This experiment showed that Duchesnea india(Audra.) Foche. has anti-tumor effect with statistical significance. This is in vitro experiment and basic experiment on Duchesnea india(Audra.) Foche. We hope more progressive researchs on Duchesnea india(Audra.) Foche. will go on and its anti-tumor effects will be more practically identified.

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Expression Levels of GABA-A Receptor Subunit Alpha 3, Gabra3 and Lipoprotein Lipase, Lpl Are Associated with the Susceptibility to Acetaminophen-Induced Hepatotoxicity

  • Kim, Minjeong;Yun, Jun-Won;Shin, Kyeho;Cho, Yejin;Yang, Mijeong;Nam, Ki Taek;Lim, Kyung-Min
    • Biomolecules & Therapeutics
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    • 제25권2호
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    • pp.112-121
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    • 2017
  • Drug-induced liver injury (DILI) is the serious and fatal drug-associated adverse effect, but its incidence is very low and individual variation in severity is substantial. Acetaminophen (APAP)-induced liver injury accounts for >50% of reported DILI cases but little is known for the cause of individual variations in the severity. Intrinsic genetic variation is considered a key element but the identity of the genes was not well-established. Here, pre-biopsy method and microarray technique was applied to uncover the key genes for APAP-induced liver injury in mice, and a cause and effect experiment employing quantitative real-time PCR was conducted to confirm the correlation between the uncovered genes and APAP-induced hepatotoxicity. We identified the innately and differentially expressed genes of mice susceptible to APAP-induced hepatotoxicity in the pre-biopsied liver tissue before APAP treatment through microarray analysis of the global gene expression profiles (Affymetrix $GeneChip^{(R)}$ Mouse Gene 1.0 ST for 28,853 genes). Expression of 16 genes including Gdap10, Lpl, Gabra3 and Ccrn4l were significantly different (t-test: FDR <10%) more than 1.5 fold in the susceptible animals than resistant. To confirm the association with the susceptibility to APAP-induced hepatotoxicity, another set of animals were measured for the expression level of selected 4 genes (higher two and lower two genes) in the liver pre-biopsy and their sensitivity to APAP-induced hepatotoxicity was evaluated by post hoc. Notably, the expressions of Gabra3 and Lpl were significantly correlated with the severity of liver injury (p<0.05) demonstrating that these genes may be linked to the susceptibility to APAP-induced hepatotoxicity.

동남아시아로부터 수입된 paradise fish Macropodus opercularis로부터 분리한 reovirus의 특성 (Characterization of an isolated reovirus from the paradise fish Macropodus opercularis imported from Southeast Asia)

  • 김위식;김수진;김정호;정성주;김도형;오명주
    • 한국어병학회지
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    • 제23권3호
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    • pp.335-342
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    • 2010
  • 2008년 3월 인도네시아로부터 수입된 paradise fish에서 대량폐사가 발생하여 병원체 검사를 실시한 결과, megalocytivirus 및 Mycobacterium sp.가 검출되었으며, CHSE-214세포에서 미동정 바이러스가 분리되었다. 본 연구에서는 미동정 바이러스의 특성을 조사하였으며, 또한 병원성 실험을 통해 paradise fish의 대량 폐사와의 연관성을 조사하였다. 분리 바이러스는 CHSE-214, BF-2, GF, SSN-1, FSP 및 FFN 세포에서 합포체의 세포변성효과를 나타내었고 IUdR 및 chloroform 처리에 감염성이 상실되지 않으며, 산 (pH 3), 알카리 (pH 11) 및 열 ($56^{\circ}C$, 30분)에 안정하였다. 핵산 분석 결과에서는 적어도 10개의 segment (0.7-3.6 kb)를 지닌 RNA 바이러스로 확인되었으며, 전자현미경 관찰 결과, 약 65 nm 크기로 인벨롭이 없으며 2중의 capsid를 지닌 바이러스가 세포질에서 관찰되었다. 이상의 결과는 Reoviridae family에 속하는 바이러스의 전형적인 특성과 일치하므로 paradise fish에서 분리된 바이러스는 reovirus로 확인되었다. Reovirus를 사용한 감염실험에서는 병원성이 나타나지 않아, reovirus는 paradise fish의 대량 폐사의 직접적인 원인이 아닐 것으로 사료되었다.

청미래덩굴 잎 물추출물이 처리된 HepG2 세포에서의 포도당흡수기전 연구 (Study of the mechanisms underlying increased glucose absorption in Smilax china L. leaf extract-treated HepG2 cells)

  • 강윤환;김대중;김경곤;이성미;최면
    • Journal of Nutrition and Health
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    • 제47권3호
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    • pp.167-175
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    • 2014
  • 본 연구에는 SCLE를 이용하여 시도된 바가 없는 glucose uptake 유도 실험을 수행하여 HepG2 세포에서 포도당흡수가 증가함을 확인하였다. 또한 이런 포도당의 흡수는 HNF-$1{\alpha}$라는 transcription factor의 활성화를 통해 GLUT-2의 발현을 증가시키기 때문인 것을 실험적으로 증명하였다. 뿐만 아니라 Bacillus stearothermophilus 유래의 GK를 이용하여 활성을 측정한 결과 SCLE가 직접적으로 GK를 활성화하여 포도당의 인산화에 영향을 주는 것을 확인할 수 있었으며 그 실험결과들은 Fig. 8에서 도식화 하였다. 본 연구를 통해 SCLE가 ${\alpha}$-glucosidase inhibition 활성에 의한 혈당의 개선과 당뇨예방 효과뿐만 아니라 다양한 세포내 기전을 통해 혈당 및 당뇨의 개선을 유도할 수 있음을 확인하였고, 이는 SCLE가 neutraceuticals 소재로서의 개발가치가 높음을 시사한다.

Association of Estrogen Receptor Alpha and Interleukin 6 Polymorphisms with Lymphovascular Invasion, Extranodal Extension, and Lower Disease-Free Survival in Thai Breast Cancer Patients

  • Sa-Nguanraksa, Doonyapat;Suntiparpluacha, Monthira;Kulprom, Anchalee;Kummalue, Tanawan;Chuangsuwanich, Tuenjai;Avirutnan, Panissadee;O-Charoenrat, Pornchai
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권6호
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    • pp.2935-2940
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    • 2016
  • Breast cancer is the most frequent type of cancer diagnosed among women worldwide and also in Thailand. Estrogen and estrogen receptors exert important roles in its genesis and progression. Several cytokines have been reported to be involved in the microenvironment that promotes distant metastasis via modulation of immune and inflammatory responses to tumor cells. Estrogen receptor genetic polymorphisms and several cytokines have been reported to be associated with breast cancer susceptibility and aggressiveness. To investigate roles of genetic polymorphisms in estrogen receptor alpha (ESR1) and interleukin 6 (IL6), breast cancer patients and control subjects were recruited from the Division of Head, Neck and Breast Surgery (Siriraj Hospital, Bangkok, Thailand). Polymorphisms in ESR1 (rs3798577) and IL6 (rs1800795 and rs1800797) were evaluated by real-time PCR in 391 breast cancer patients and 79 healthy controls. Associations between genetic polymorphisms and clinicopathological data were determined. There was no association between genetic polymorphisms and breast cancer susceptibility. However the ESR1 rs3798577 CT genotype was associated with presence of lymphovascular invasion (OR=2.07, 95%CI 1.20-3.56, p=0.009) when compared to the TT genotype. IL6 rs1800795 CC genotype was associated with presence of extranodal extension (OR= 2.30, 95%CI 1.23-4.31, p=0.009) when compared to the GG genotype. Survival analysis showed that IL6 rs1800797 AG or AA genotypes were associated with lower disease-free survival. These findings indicate that polymorphisms in ESR1 and IL6 contribute to aggressiveness of breast cancer and may be used to identify high risk patients.

조직공학적 연골재생을 위한 In Vitro 환경에서의 탈미네랄화 골분용액을 함유한 PLGA 지지체의 효과 (Effect of PLGA Scaffold Containing Demineralized Bone Solution for Articular Cartilage Tissue Engineering: In Vitro Test)

  • 안우영;김혜린;송정은;이동원;강길선
    • 폴리머
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    • 제35권6호
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    • pp.499-504
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    • 2011
  • 본 연구팀은 탈미네랄화 골분(DBP)이라는 천연재료를 졸(sol)화시켜 poly(lactide-co-glycolide) (PLGA)에 함침시킨 지지체를 개발하였다. DBP를 함침시킨 PLGA 지지체 상의 세포증식과 모폴로지를 평가하기 위해 MTT 분석과 SEM을 측정하였다. 또한 sGAG와 콜라겐 함량 측정과 파종된 연골 세포의 표현형 유지에 미치는 영향을 확인하였다. 그 결과 PLGA에 DBP를 함침시킨 지지체가 PLGA 지지체보다 높은 세포 증식률을 보였다. 또한 파종된 연골세포의 표현형 유지에도 긍정적인 영향을 미치는 것을 확인하였다. 이번 연구 결과를 토대로 PLGA에 DBP를 이용한 용액을 함침시킴으로써 DBP내의 성장인자와의 상호작용을 통해 연골세포의 성장에 긍정적 영향을 미쳐 안정되게 연골을 조직화할 수 있는 연골조직공학 지지체로 적합할 것으로 예상된다.

공동배양과정의 배지조성과 배양조건이 벼 형질전환효율에 미치는 영향 (The Effects of Co-cultivation Medium and Culture Conditions on Rice Transformation Efficiency)

  • 김율호;박향미;최만수;윤홍태;최임수;신동범;김정곤;이장용
    • 한국육종학회지
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    • 제41권3호
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    • pp.252-260
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    • 2009
  • 1. 본 연구에서는 공동배양 배지에 Agrobacterium 성장 억제물질인 silver nitrate를 첨가하고 변온과 여과지처리를 추가하여 공동배양 기간을 7일로 늘였으며, 또한 항산화 물질 3종을 공동배양 배지에 첨가하여 세포의 oxidative burst를 최소화함으로써 벼 형질전환효율을 높일 수 있었다. 또한 이 방법을 적용하여 형질전환이 어려운 품종을 대상으로도 형질전환 식물체를 작성할 수 있었다. 2. 벼 형질전환체의 70%에서 도입유전자 수가 1copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입됨을 확인 하였다. 3. 이러한 결과를 바탕으로, 새로운 공동배양 방법을 사용하여 우수한 농업적 형질을 가진 벼 육종 소재 및 품종을 신속하게 개발할 수 있을 것으로 기대된다.

Breeding on High Lycopene and Beta Carotene with Multi-Disease Resistance in Tomato

  • Kim, Myung Kwon;Lee, Hee Bong
    • 한국육종학회지
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    • 제41권1호
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    • pp.1-8
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    • 2009
  • This study was carried out to breed and develop high quality and functional nutrient tomato with multi disease resistance as well as a stable growing adaptation for fresh market usage under protected plastic houses cultivation. The materials were used 5 inbred lines and their 6 hybrids of large tomato group, which have been bred and developed from 1999 to 2007 in Division of Plant Resource Department of Chungnam National University. Fruit weight showed hybrid vigor effect that $F_1$ hybrids weighed more than their parent lines, fruit shape formed three type of oblate, deep oblate and globe shape, in firmness and pericarp thickness have got a high significant correlation, inbred DN611 line was measured the most firm fruit with 6.04 mm pericarp thickness. In fruit color at maturity, pink color crossed to red color appeared all red fruit color in the $F_1$ hybrids, it means red skin color is a dominant gene compared to pink skin color is a recessive gene in tomato, while between fruit skin color and shoulder part color showed no any co-relationship. The sugar content and titratable acid of $F_1$ hybrids inherited an intermediate data of their parent lines, the flavor of KP543 inbred line and the hybrid (JB535 x KP543) revealed the better taste with high brix and proper titratable acid content$^{*}$. In beta carotene content DN611 line showed 2~3 times higher than other materials so that its 3 hybrids contained an increased level of beta carotene, lycopene content was not so much difference among inbred lines and $F_1$ hybrids, of them MD508 contained higher of 8.72 mg and hybrid (JB535 x JA517) had 8.05 mg lycopene content per 100 g fruit, overall pink skin color and red skin color measured a higher lycopene content than yellow and orange skin color at ripe stage. In disease resistance test by PCR marker for Fusarium race2 (I2), Nematode (Mi1), ToMV ($Tm2^2$), Cladosporium (Cf9), (JB535 x JA517) hybrid have got multi-resistance with homozygote band in Nematode, ToMV, Cladosporium and heterozygote band in Fusarium race2. Through this breeding program we could select high quality and functional nutrient with multi resistant $F_1$ hybrids and inbred lines in tomato which are two best hybrids (JB535 x MD508), (JB535 x JA517), additionally developed high beta carotene inbred line DN611 and increased the level of lycopene inbred line MD508. These results will be very useful to make a high quality tomato variety continuously.