• 생명과학회지
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• 제25권4호
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• pp.376-384
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• 2015

소의 질을 평가하기 위해서는 중요한 인자인 근육내 지방(또는 마블링)을 조절하는 분자를 연구해야 한다. cDNA microarray를 사용하여 등지방 조직과 최장근의 유전자발현 차이를 비교하였다. 이 연구를 통해, 우리는 한우의 지방조직에 1211개, 근육조직에서 1346개의 특이 유전자를 확인하였다. bovine chip은 지방조직의 920개 유전자와 근육조직의 760개 유전자로 이루어진 1680개의 특이 유전자로 구성되어있다. 이 실험에서 Microarray 분석은 등지방조직(Cy3)과 최장근(Cy5)의 유전자 발현에 있어서 큰 차이를 보여준다. 차이를 보이는 많은 특이유전자 중에서, 12-리폭시게나아제 유전자와 프로스타글란딘 D 합성효소는 근육내 지방의 축적을 조절하는 중요한 효소이다. 본 연구에서, 일반적으로 발현되지만 한우의 근육과 지방 조직에서 차이를 보이는 많은 유전자를 hybridization 분석을 통해 발견하였다. 선택된 유전자의 발현 수준은 반정량적 RT-PCR을 통해 확인하였고, 그 결과는 cDNA microarray와 유사하였다.

• Journal of Microbiology and Biotechnology
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• 제24권3호
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• pp.363-370
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• 2014

Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.

• 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.

• Journal of Animal Science and Technology
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• 제47권1호
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• pp.1-10
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• 2005

Intramuscular fat content(lMF) is considered as one of major economic traits in the pig breeding and industry. In general, high IMF results in better meat quality. Several approaches to detect quantitative trait 10ci( QTL) for IMF indicated a strong possibility of the existence of a QTL related to IMF between the microsatellite marker SW71 and SW1881 on SSC6q. Porcine FABP3 has been considered as a candidate gene affecting IMF due to its physiological roles and position on the pig genome. Two novel mutations, g.-114T> C and g.-158T>G were detected by duplicate sequencing of the porcine FABP3 promoter region. These two mutations were identified as absolute linkage disequilibrium. The g.-158T> G mutation was used for investigating relationships with growth and fat deposition traits. The GG genotype of the g.-158T> G polymorphism showed highly negative effects(P< 0.01) on body weights at 3 and 12 weeks of age, and a positive effect(P< 0.05) on IMF. However, backfat thickness(BF) and carcass fat(CF) content were not significantly associated with the genotype. The result indicates that the novel mutations, identified in this study, could be utilized as possible genetic markers to improve IMF, independent with BF.

• 대한예방한의학회지
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• 제16권3호
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• pp.139-153
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• 2012

Objectives : NK cells are spontaneously cytotoxic lymphocytes. These are not only important parts in the first line of defence against bacterial and viral infections of outside, but they may also play a critical role in chronic viral diseases. NK cells kill their targets spontaneously, without the need for prior sensitization and class I MHC restriction by the regulation of cytolytic functions and secretion of a variety of cytokines, such as interleukin-12(IL-12), MCP-1, IL-6, TNF-${\alpha}$, IFN-${\gamma}$. In addition, macrophage and NK cells cooperate through the production of cell mediates. These cooperation and modulation are one of major factors to prevent for evading immune surveillance of cancer. Hence, it could be assumed that if any candidate to enhance activities of macrophage and NK cell, it is considered as a potentially useful agents against cancer. Methods : In our study, to investigate effect of fermented soybean extracts by Lactic acid bacteria (SFE, soybean fermented extracts) work on intestinal immune cell to maintain general immune modulating and anti-cancer activity. We analyzed NK cytotoxicity assay and gene expressions of cytokine related with macrophage and NK cell activity. Results : In vitro experiment, SFE was verified as safety material for cell toxicicty to tumor cell strain without any toxicity of tumor growth inhibition and various cell strain. Effects of macrophage activity stimulating directly by SFE measured induced cytokine. The studies showed that IL-12 production by stimulation of SFE depended on concentration from 0.16mg/mL to 0.63mg/mL with non toxicity to cell, and it was the best activity at 0.63mg/mL. Besides, the effective concentration of SFE producing TNF-${\alpha}$ is similar to IL-12, but it was the best activity at 1.25mg/mL. The level of MCP-1, IL-6 and IFN-${\gamma}$ depended on concentration from 0.16mg/mL to 10mg/mL, IFN-${\gamma}$ showed the best activity at the effective concentration of 0.63mg/mL. With the result of NK cell activity measurement, the spleen cell of mouse injected SFE had 1.5 times higher killing effect than non injected cell. Conclusions : The result of this studies is that Soybean fermetated extracts(SFE) has possibility to immune aided material for the function not only inhibition of microbial infection to macrophage but also activity of adaption immune and cellular immune system.

• Genes and Genomics
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• 제40권11호
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• pp.1169-1180
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• 2018

Cave shrimps from the genera Typhlatya, Stygiocaris and Typhlopatsa (TST complex) comprises twenty cave-adapted taxa, which mainly occur in the anchialine environment. Anchialine habitats may undergo drastic environmental fluctuations, including spatial and temporal changes in salinity, temperature, and dissolved oxygen content. Previous studies of crustaceans from anchialine caves suggest that they have possessed morphological, behavioral, and physiological adaptations to cope with the extreme conditions, similar to other cave-dwelling crustaceans. However, the genetic basis has not been thoroughly explored in crustaceans from anchialine habitats, which can experience hypoxic regimes. To test whether the TST shrimp-complex hypoxia adaptations matched adaptive evolution of mitochondrial OXPHOS genes. The 13 OXPHOS genes from mitochondrial genomes of 98 shrimps and 1 outgroup were examined. For each of these genes was investigated and compared to orthologous sequences using both gene (i.e. branch-site and Datamonkey) and protein (i.e. TreeSAAP) level approaches. Positive selection was detected in 11 of the 13 candidate genes, and the radical amino acid changes sites scattered throughout the entire TST complex phylogeny. Additionally, a series of parallel/convergent amino acid substitutions were identified in mitochondrial OXPHOS genes of TST complex shrimps, which reflect functional convergence or similar genetic mechanisms of cave adaptation. The extensive occurrence of positive selection is suggestive of their essential role in adaptation to hypoxic anchialine environment, and further implying that TST complex shrimps might have acquired a finely capacity for energy metabolism. These results provided some new insights into the genetic basis of anchialine hypoxia adaptation.

• Asian-Australasian Journal of Animal Sciences
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• 제32권9호
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• pp.1458-1468
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• 2019

Objective: As one of the most important metabolic organs, the liver plays vital roles in modulating the lipid metabolism. This study was to compare miRNA expression profiles of the Large White liver between two different developmental periods and to identify candidate miRNAs for lipid metabolism. Methods: Eight liver samples were collected from White Large of 70-day fetus (P70) and of 70-day piglets (D70) (with 4 biological repeats at each development period) to construct sRNA libraries. Then the eight prepared sRNA libraries were sequenced using Illumina next-generation sequencing technology on HiSeq 2500 platform. Results: As a result, we obtained 346 known and 187 novel miRNAs. Compared with the D70, 55 down- and 61 up-regulated miRNAs were shown to be significantly differentially expressed (DE). Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis indicated that these DE miRNAs were mainly involved in growth, development and diverse metabolic processes. They were predicted to regulate lipid metabolism through adipocytokine signaling pathway, mitogen-activated protein kinase, AMP-activated protein kinase, cyclic adenosine monophosphate, phosphatidylinositol 3 kinase/protein kinase B, and Notch signaling pathway. The four most abundantly expressed miRNAs were miR-122, miR-26a and miR-30a-5p (miR-122 only in P70), which play important roles in lipid metabolism. Integration analysis (details of mRNAs sequencing data were shown in another unpublished paper) revealed that many target genes of the DE miRNAs (miR-181b, miR-145-5p, miR-199a-5p, and miR-98) might be critical regulators in lipid metabolic process, including acyl-CoA synthetase long chain family member 4, ATP-binding casette A4, and stearyl-CoA desaturase. Thus, these miRNAs were the promising candidates for lipid metabolism. Conclusion: Our study provides the main differences in the Large White at miRNA level between two different developmental stages. It supplies a valuable database for the further function and mechanism elucidation of miRNAs in porcine liver development and lipid metabolism.

• 생명과학회지
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• 제29권9호
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• pp.1016-1022
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• 2019

고령화 사회에서 비만을 예방하는 것에 대한 관심이 높아지는 추세에 따라, 남성과 여성 모두 비만 관리에 상당한 비용과 노력을 지불하고 있는 실정이다. 본 연구에서는 항비만 효과를 증명하기 위해서는 3T3-L1 지방전구세포를 이용한 연구가 필수적으로 수행되기 때문에, 3T3-L1 지방전구세포에서 항비만 효과를 지닌 천연물에 조사하였다. 70% 에탄올을 이용한 사상자(Torilis Japonica Decandolle) 추출물이 3T3-L1 지방전구세포에서 성숙한 지방세포로의 분화에 미치는 효과를 Oil red O assay, western blot을 통해 확인하였다. 대조군에 비해 사상자 추출물의 $100{\mu}g/ml$ 농도에서 지방세포 분화와 세포 내 triglyceride (TG) 수준을 효과적으로 억제하였다. TG 함량 감소 메커니즘을 규명하기 위해, peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) 및 CCAAT enhancer-binding-proteins-${\alpha}$ ($C/EBP{\alpha}$), acetyl-CoA carboxylase (ACC)의 인산화 등 항비만 관련 단백질의 표현 수준을 확인하였다. 그 결과, 사상자 추출물은 $PPAR{\gamma}$, $C/EBP{\alpha}$, ACC 인산화 단백질의 발현 정도를 현저하게 감소시켰다. 종합하자면, 사상자 추출물이 비만 예방에 가장 효과적인 후보임을 명백하게 보여준다. 더 나아가서, 천연물인 사상자의 항비만 효과에 핵심적인 역할을 수행하는 활성 화합물을 탐색 및 분리하기 위한 추가 연구가 필요하다고 사료된다.

• 생명과학회지
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• 제31권2호
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• pp.175-182
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• 2021

기능성 화장품 소재로서 활용할 수 있는 효모의 분리를 위하여 순창군 베리류 및 과수원 토양에서 분리주 80종을 1차로 선별하였다. 80종의 분리주를 대상으로 항산화 활성 및 tyrosinase 저해 활성을 측정한 결과 DPPH 라디칼 소거능은 51.41%, SOD 활성은 62.23%, tyrosinase 저해 활성 64.75%로 가장 우수한 FT4-4를 최종적으로 선별하였다. 18S rRNA 염기서열 분석을 통해 Saccharomyces cerevisiae FT4-4로 명명하였으며, API ZYM을 이용하여 세포 외 효소 활성을 추가로 측정하였다. 발효 시간에 따른 균체 성장 및 tyrosinase 저해 활성의 변화를 측정한 결과 배양 후 16시간에 최대 균체량인 3.16 g/l와 67.68%의 tyrosinase 저해활성을 나타내었다. 또한, S. cerevisiae FT4-4의 화장품 소재로 활용하기 위한 세포 독성과 melanoma B16F10 세포 멜라닌 억제능을 확인한 결과, 세포독성은 50 mg/ml 이하의 농도에서 100% 이상의 세포 생존율을 보였으며, 시료 10 mg/ml에서의 멜라닌 생합성 저해능은 72.02%로 측정되었다. 향후 FT4-4의 화장품 소재로 활용하기 위해서는 생산 수율을 증가하기 위한 생산조건 확립 이외에도 안전성을 향상시키기 위한 추가적인 독성연구 등 다양한 연구가 수반되어야 하나, 본 연구에서의 항산화 및 미백 효능만으로도 충분히 활용할 가치가 있는 소재로 사료된다.

• 대한본초학회지
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• 제37권3호
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• pp.9-19
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• 2022

Objectives : Angelicae Gigantis Radix (AG) is a plant of the Ranunculus family. AG have been reported to have various pharmacological effects on human health which include uterine growth promotion, anti-inflammatory, analgesic, and immune enhancement. However, research on dermatitis disease is insufficient. Therefore, we investigated the effects of AG on tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) stimulated HaCaT cell. Methods : To investigate the effect of AG on HaCaT cell, HaCaT cells were pre-treated with AG for 1 hour and then stimulated with TNF-α/IFN-γ. After 24 hours, media and cells were harvested to analyze the inflammatory mediators. Concentration of human interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α in the media were assessed by ELISA. mRNA expression of human thymus and activation-regulated chemokine (TARC), IL-6, and IL-8 were analyzed by RT-PCR. Additionally, the mechanisms of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway were investigated by Western blot. Results : The treatment of AG inhibited gene expression levels of IL-6, IL-8, and TARC and protein expression levels of IL-1β, MCP-1, and GM-CSF. Also, AG significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation and NF-κB translocation in TNF-α/IFN-γ stimulated HaCaT cell. Conclusions : Taken together, these results demonstrate that AG can alleviate inflammatory diseases such as atopic dermatitis by regulating the expression of inflammatory cytokines. Also, it suggest that AG may a promising candidate drug for the treatment of inflammatory disease such as atopic dermatitis.