• 대한한의학방제학회지
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• 제28권3호
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• pp.255-269
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• 2020

Objectives : Hemistepta lyrata Bunge (Bunge) is a wild herb that has been used for managing fever and wound in Korean Traditional Medicine. The present study explored the effects of H. lyrata extract on liver X receptor (LXR) α-dependent lipogenic genes in hepatocyte-derived cells. Methods : After HepG2 cells or Huh7 cells were pre-treated with 1-10 ㎍/mL of H. lyrata extract or its fractionated extract for 0.5 h, the cells were subsequently exposed to LXR ligand for 6-24 h. Cell viability, LXR response element (LXRE)-driven luciferase activity, sterol regulatory element binding protein-response element (SREBP-RE)-driven luciferase activity, SREBP-1c expression, and mRNA levels of LXRα and its-dependent target genes were determined. In addition, LC-MS/MS analysis was conducted to explore major compounds in H. lyrata-chloroform fractionated extract #4 (HL-CF4). Results : Of various H. lyrata extracts tested, chloroform extract and its fractionated extract #4, HL-CF4, significantly decreased T0901317-mediated SREBP-1c expression. In addition, HL-CF4 significantly reduced LXRE atransactivation and LXRα mRNA expression without any cytotoxicity. Moreover, HL-CF4 prevented the SREBP-RE-driven luciferase activity and mRNA levels of fatty acid synthase and stearoyl-CoA desaturase-1 induced by T0901317. Results from LC-MS/MS analysis at positive/negative mode indicated that HL-CF4 contained several compounds showing m/z 197.1176 (C₁₁H₁₇O₃), 693.2913/227.1069 (C₃₈H₄₅O₁₂/C₁₅H₁₅O₂), 203.1797 (C₁₅H₂₃), 181.1225 (C₁₁H₁₇O₂), 591.2957 (C₃₅H₄₃O₈), 379.1040 (C₁₈H₁₉O₉), 409.1509 (C₂₀H₂₅O₉), 309.1348 (C₁₆H₂₁O₆), 391.1404 (C₂₀H₂₃O₈), and 669.2924/389.1248 (C₃₆H₄₅O₁₂/C₂₀H₂₁O₈). Conclusion : Based on its inhibition of the LXRα-dependent signaling pathway, H. lyrata chloroform extract and HL-CF4 have prophylactic potentials for managing non-alcoholic fatty liver.

• The Plant Pathology Journal
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• 제15권3호
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• pp.137-143
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• 1999

A novel chromosomal gene tagging technique using a specific fragment of the fatty acid desaturase-like open reading frame (des-like ORF) from the tox-argK gene cluster of Pseudomonas syringae pv. phaseolicola was developed to identify Xanthomonas spp.released into the environment as biocontrol agents. X. campestris pv. convolvuli FB-635, a pathogen of Convolvulus arvensis L., (bindweed), was chosen as the organism in which to develop and test the system. A 0.52 kb DES fragment amplified from P. syringae pv. phaseolicola C-199 was inserted into pGX15, a cosmid clone containing a 10.3 kb Eco RI-HindIII fragment derived from the xanthomonadin biosynthetic gene cluster contained in plasmid pIG102, to create a pigG::DES insertion. The 10.8 kb EcoRI-BamHI fragment carrying the pigG:: DES insertion was cloned into pLAFR3 to generate pLXP22. pLXP22 was then conjugated into X. campestris pv. convolvuli FB-635 and the pigG::DES insertion integrated into the bacterial chromosome by marker exchange. Rifampicin resistant, tetracycline sensitive, starch hydrolyzing, white colonies were used to differentiate the marked strain from yellow pigmented wild-type ones. PCR primers specific for the unique DES fragment were used for direct detection of the marked strain. Result showed the marked strain could be detected at very low levels even in the presence of high levels of other closely related or competitive bacteria. This PCR-based DES-tagging system provides a rapid and specific tool for directly monitoring the dispersal and persistence of Xanthomonas spp.released into the environment.

• Asian-Australasian Journal of Animal Sciences
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• 제27권12호
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• pp.1783-1793
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• 2014

Capsaicin is a major constituent of hot chili peppers that influences lipid metabolism in animals. In this study, we explored the effects of capsaicin on adipogenic differentiation of bovine bone marrow mesenchymal stem cells (BMSCs) in a dose- and time-dependent manner. The BMSCs were treated with various concentrations of capsaicin (0, 0.1, 1, 5, and $10{\mu}M$) for 2, 4, and 6 days. Capsaicin suppressed fat deposition significantly during adipogenic differentiation. Peroxisome proliferator-activated receptor gamma, cytosine-cytosine-adenosine-adenosine-thymidine/enhancer binding protein alpha, fatty acid binding protein 4, and stearoyl-CoA desaturase expression decreased after capsaicin treatment. We showed that the number of apoptotic cells increased in dose- and time-dependent manners. Furthermore, we found that capsaicin increased the expression levels of apoptotic genes, such as B-cell lymphoma 2-associated X protein and caspase 3. Overall, capsaicin inhibits fat deposition by triggering apoptosis.

• 한국미생물·생명공학회지
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• 제49권2호
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• pp.210-216
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• 2021

본 연구에서는 CRISPR/CAS9을 이용하여 S. cerevisiae의 ACC1, ELO1, OLE1 유전자의 프로모터를 TEF1으로 교체하여 그 발현량을 증가시키고 그에 따른 에탄올에 대한 저항성과 생산량 변화를 확인하였다. 18% 에탄올이 함유된 YPD 배지에서 control을 제외하고 유전자 과발현을 일으킨 mutant 균주 모두가 24시간까지 viable하게 생존하는 것을 확인하였다. 에탄올 발효에서는 유전자 과발현 균주 모두가 에탄올 수율에서 ACC1 과발현 균주가 428.18 ± 0.29 mg/g, ELO1 과발현 균주는 416.15 ± 4.3 mg/g, OLE1 과발현 균주는 430.55 ± 6.00 mg/g에 도달하였으며, 이는 control의 수율인 400.26 ± 0.42 mg/g 보다 높은 수준에 도달하였다. 이 결과는 높은 농도의 에탄올에서 탄소 사슬이 긴 불포화지방산의 비율이 증가한다는 연구결과가 역 또한 성립한다는 것을 증명하였다. ELO1의 과발현은 elongation of fatty acid protein의 생산 증가를 불러 일으킨다. 또한 OLE1도 acylCoA desaturase 효소의 활성을 증대시킨다. TEF1이라는 strong promoter를 이용한 이번 실험에서 ELO1 과발현 균주가 OLE1 과발현 균주보다 S. cerevisiae의 에탄올 저해 감소와 발효에 긍정적인 영향을 미침을 확인하였다.

• Journal of Ginseng Research
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• 제43권4호
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• pp.572-579
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• 2019

Background: Panax ginseng has been used in traditional medicine to strengthen the body and mental well-being of humans for thousands of years. Many elite ginseng cultivars have been developed, and ginseng cultivation has become well established during the last century. However, heat stress poses an important threat to the growth and sustainable production of ginseng. Efforts have been made to study the effects of high temperature on ginseng physiology, but knowledge of the molecular responses to heat stress is still limited. Methods: We sequenced the transcriptomes (RNA-Seq) of two ginseng cultivars, Chunpoong (CP) and Yunpoong (YP), which are sensitive and resistant to heat stress, respectively, after 1- and 3-week heat treatments. Differential gene expression and gene ontology enrichment along with profiled chlorophyll contents were performed. Results: CP is more sensitive to heat stress than YP and exhibited a lower chlorophyll content than YP. Moreover, heat stress reduced the chlorophyll content more rapidly in CP than in YP. A total of 329 heat-responsive genes were identified. Intriguingly, genes encoding chlorophyll a/b-binding proteins, WRKY transcription factors, and fatty acid desaturase were predominantly responsive during heat stress and appeared to regulate photosynthesis. In addition, a genome-wide scan of photosynthetic and sugar metabolic genes revealed reduced transcription levels for ribulose 1,5-bisphosphate carboxylase/oxygenase under heat stress, especially in CP, possibly attributable to elevated levels of soluble sugars. Conclusion: Our comprehensive genomic analysis reveals candidate loci/gene targets for breeding and functional studies related to developing high temperature-tolerant ginseng varieties.

• Ocean and Polar Research
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• 제44권1호
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• pp.61-67
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• 2022

In this study, in order to evaluate the anti-obesity effect of sargassum horneri extract, the effects of the extract on lipase activity and preadipocyte differentiation in 3T3-L1 cells were investigated. S. horneri extract between 0.0 and 1.0 mg/mL showed no cytotoxicity and inhibited lipase activity by 68.1%. When S. horneri extract was utilized at levels of 0.25, 0.5, and 1.0 mg/mL in 3T3-L1 cells, preadipocytes differentiation decreased by 11.4, 19.7, and 25.6%, respectively, showing anti-obesity effects. In addition, after treatment with 1.0 mg/mL S. horneri extract, the mRNA expression levels of sterol regulatory element binding proteins-1c (SREBP-1c), peroxisome proliferator activated receptor-γ (PPAR-γ), CCAAT enhancer binding protein-α (CEBP-α), fatty acid synthase (FAS), and stearoyl-CoA desaturase1 (SCD1) in 3T3-L1 cells were significantly decreased (p < 0.05) by 65.2, 54.9, 50.0, 33.8, and 33.8% respectively. These results showed that S. horneri extract suppresses lipase activity and prophylactic preadipocyte differentiation in 3T3-L1, and thus can be used as an anti-obesity agent in functional foods and medicines.

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1173-1183
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• 2022

Adipogenesis is a complex process comprising commitment and a differentiation stages. Through research, many different transcriptional factors were found to mediate preadipocyte commitment and differentiation. Lysine has a potential of regulating the commitment and differentiation of preadipocytes. In the present study, intramuscular stromal vascular cells (SVC) isolated from Hanwoo beef cattle were used to elucidate the effects of low lysine level on adipogenesis. SVC were isolated and incubated with various concentrations of lysine (0, 37.5, 75, 150 and 300 µg/mL). No significant difference were observed in the proliferation of SVC after 24 and 48 h of incubation with different concentration of lysine. On preadipocyte determination, reducing the level of lysine significantly increased the expression of preadipocyte commitment gene Zinc finger protein 423 and Preadipocyte factor-1. Upon differentiation, Oil Red O staining revealed that lipid accumulation and triglyceride content significantly increased with the decreasing lysine levels in the media. Expression levels of peroxisome proliferator-activated receptor-γ, CCAAT enhancer binding protein-α, sterol regulatory element binding protein-1c, Fatty Acid Binding Protein 4 and stearoyl CoA desaturase were upregulated by the decreased level of lysine. These data suggest the potential mechanism of action for the improved preadipocyte commitment and adipocyte differentiation in bovine intramuscular SVC upon treatment with low levels of lysine. These findings may be valuable in developing feed rations that promote deposition of intramuscular fat in beef cattle through lysine level modification.

• 한국식품영양과학회지
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• 제45권6호
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• pp.789-796
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• 2016

본 연구에서 고당식이로 비알코올성 지방간을 유도한 마우스의 체중 변화는 대조군보다 백미군, 혼합잡곡군, 항비만혼합잡곡군에서 체중증가율이 낮았고 간 무게 또한 유의적으로 감소했으며, 간 내 조직학적 지방구 수와 크기가 감소한 것을 관찰할 수 있었다. 혈청 지질 수치 역시 개선 효과를 보였는데 모든 실험군이 대조군보다 중성지방, 총콜레스테롤 및 저밀도 콜레스테롤의 농도가 감소하였고, 혈청 고밀도 콜레스테롤은 모두 증가하였다. 간 조직 내 지질합성 및 지방산 침투와 관련 유전자 인자에서 대조군보다 SREBP-1c mRNA 유전자 발현 수준은 백미군, 혼합잡곡군 및 항비만혼합잡곡군에서, ACC 및 FAS mRNA 유전자 발현 수준은 혼합잡곡군과 항비만혼합잡곡군에서, SCD-1 mRNA 유전자 발현 수준은 항비만혼합잡곡군에서 감소하였다. CD36 및 $PPAR-{\gamma}$ mRNA 유전자 발현 수준 또한 대조군보다 백미군, 혼합잡곡군, 항비만혼합잡곡군에서 감소하였다. 간 내 ${\beta}$산화로 지방축적 억제와 관련된 유전자 인자인 $PPAR-{\alpha}$ 및 CPT-1 mRNA 유전자 발현 수준은 대조군보다 혼합잡곡군, 항비만혼합잡곡군에서 증가하였다. 본 실험 결과를 종합해 볼 때 고당식이로 비알코올성 지방간질환을 유도한 마우스에서 백미군, 혼합잡곡군 및 항비만혼합잡곡군 모두 지질 대사 개선 효과가 나타났으며 항비만혼합잡곡군이 가장 효과적이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권5호
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• pp.621-628
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• 2012

The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

• 한방비만학회지
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• 제15권2호
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• pp.75-92
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• 2015

Objectives: This study was designed to investigate the effect of Gami-cheongpyesagan-tang extract (GCST) on high fat diet-induced obesity in rats. Methods: The mice were divided into six groups; normal diet control, high fat diet control (HFD), HFD+GCST administrated group (100, 200, and 400 mg/kg) and olistat-admistrated group. Obesity was induced by high fat diet (45%) for 7 weeks in mice, and GCST was administrated orally every day for 7 weeks. The body weight, food intake, and serological markers such as total cholesterol, triglyceride, lipid contents, leptin, adiponectin and glutamic oxaloacetic transaminase/glutamic pyruvic transaminase were measured in mice. The mRNA expression of obese-associating genes such as sterol regulatory element-binding protein (SREBP)-1c, fatty acid synthase (FAS), stearoyl-CaP desaturase (SCD-1), peroxisome proliferator-activated receptor $(PPAR)-{\alpha}$, COA oxidase (ACO), and carnitine palmitoyltransferase ($CPT-1{\alpha}$) was analyzed by reverse transcription polymerase chain reaction. Results: The administration of GCST at 400 mg/kg, significantly reduced the increase of body weight and food intake as well as food efficiency compared to HFD group. GCST decreased the serum levels of triglyceride, total cholesterol, low-density lipoprotein-cholesterol, leptin in HFD control group and inhibited lipid accumulation in liver and adipose tissues, but did not increase high-density lipoprotein-cholesterol. In the liver tissues of GCST administrated HFD group, the mRNA levels of SREBP-1c, FAS and SCD-1 were decreased and the mRNA levels of $PPAR-{\alpha}$, ACO, and $CPT-1{\alpha}$ were increased. Conclusions: These results indicate that GCST could improve high fat diet induced obesity through inhibiting the hyperlipidemia in fatty Liver. It suggest that GCST may be used clinically for declining the accumultion of body fat with hyperlipidemia.