• Animal Bioscience
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• 제35권5호
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• pp.763-777
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• 2022

Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

• 한방재활의학과학회지
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• 제32권4호
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• pp.1-8
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• 2022

Objectives This study is to investigate the effects and mechanisms of Gastrodia elata extract (GEE) on the high-fat diet-induced obesity model. Methods C57BL/6 mice were randomly assigned into 5 groups (n=10). Control group was fed normal diet (ND). Obesity group was fed 60% high fat diet (HFD). The other three groups were fed HFD with 100, 200, 500 mg/kg GEE. After five weeks, body weight, liver and epididymal fat weight, triglyceride concentration in liver and serum, sterol regulatory element-binding protein-1 (SREBP-1), acetyl-CoA carboxylase (ACC), fatty acid synthase, peroxisome proliferator-activated receptor 𝛾 (PPAR-𝛾), CCAAT/enhancer binding protein 𝛼 (C/EBP-𝛼) expression level, insulin concentration in serum were measured. Results The GEE (100, 200, and 500 mg/kg)-treated animals exhibited substantial decreases in body mass, liver weight and epididymal white adipose tissue collate to the HFD-fed group. GEE treatment also reduced hepatic and serum triglyceride level. Furthermore, GEE treatment significantly inhibited adipogenesis in the GEE group by reducing the protein expression of SREBP-1, ACC and the messenger RNA expression of PPAR𝛾, C/EBP-𝛼, which are adipocyte differentiation-related genes. Conclusions These research outcomes recommend that GEE is possibly valuable for the prevention of HFD-induced obesity via modification of various pathways related with adipogenesis and adipocyte differentiation.

• Animal Bioscience
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• 제37권5호
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• pp.929-943
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• 2024

Objective: The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

• 한국식품과학회지
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• 제51권3호
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• pp.272-277
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• 2019

본 연구진은 선행연구에서 MCE-DAG를 섭취한 마우스에서 혈중 총 콜레스테롤과 LDL 콜레스테롤의 감소를 보고한 바 있어, 본 연구에서 in vitro를 통해 MCE-DAG와 간의 콜레스테롤 항상성 기전의 관련성을 구명하고자 하였다. LDLR과 같은 콜레스테롤 흡수 관련 인자의 발현이 MCE-DAG에 의해 증가한 반면, LDLR을 억제하는 PCSK9의 발현은 감소하였다. 또한, 콜레스테롤 합성 관련 인자인 HMGCR의 발현이 MCE-DAG에 의해 증가하였고, 전사조절인자인 SREBP2의 발현이 증가하였다. 이러한 결과들은 콜레스테롤의 합성과 흡수가 동시에 증가하였음을 뒷받침한다. 즉, 간 내 콜레스테롤 필요량이 증가함에 따라, 간의 콜레스테롤 합성 및 흡수를 활성화시켜 콜레스테롤 항상성을 유지하는 기전이 촉진되었음을 의미한다. 하지만 간 세포 내 총 콜레스테롤 양은 MCE-DAG에서 영향을 받지 않았다. 콜레스테롤 흡수 및 합성 기전이 촉진되었음에도 세포 내 콜레스테롤 농도가 증가하지 않은 현상은 담즙산 등 콜레스테롤 분비 촉진에 의한 것일 수 있다. 이러한 추론은 추후 콜레스테롤 분비 기전을 검증할 수 있는 실험을 설계하여 검증해볼 필요성이 있다. 결론적으로 MCE-DAG는 세포 내 콜레스테롤 흡수 작용을 촉진하는 효과가 있어 추후 기능성 유지로 활용 가능할 것으로 판단된다.

• 대한약침학회지
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• 제9권2호
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• pp.5-16
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• 2006

Objective : It has long been known about the osteogenic effect of CPC-HAS on bone tissues. However, it has not been determined the effect of CPC-HAS on cancer cells. The purpose of this study is to screen the CPC-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells. Oligonucleotide microarray and proteomics approaches were employed to screen the differential expression genes. Methods : CPC-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CPC-HAS (0.1, 0.5, 1.5, 10, 20mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cell in all concentrations(0.l, 0.5, 1.5, 10, 20mg/ml). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 23 with 5 up-regulated and 18 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. Two down-regulated proteins were aldehyde dehydrogenase 1 and enolase 1, and up-regulated protein was fatty acid binding protein 1 by 1.5mg/ml of CPC-HAS. Discussion : This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray and proteomic analysis. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.

• Journal of Animal Science and Technology
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• 제63권4호
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• pp.934-953
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• 2021

Ciglitazone is a member of the thiazolidinedione family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ), thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC). Muscle-derived BSCs were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 µM ciglitazone (CL), 10 µM ciglitazone (CM), or 20 µM ciglitazone (CH). Control (CON) BSC were cultured only in a differentiation medium (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 (Pax7) proteins was confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase, stearoyl-CoA desaturase, and perilipin 2. Ciglitazone treatments increased gene expression of Pax3 and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5, and myogenin-4 (p < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (p < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that DEGs were involved in the p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, tumor necrosis factor signaling pathway, non-alcoholic fatty liver disease, PI3K-Akt signaling pathway, and Wnt signaling pathway. These results indicate that ciglitazone acts as PPARγ agonist, effectively increases the adiponectin concentration and adipogenic gene expression, and stimulates the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.

• 한국식품저장유통학회지
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• 제22권5호
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• pp.744-750
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• 2015

본 연구에서는 연화 열수 및 에탄올 추출물의 3T3-L1 지방세포 분화 및 ROS생성 저감효과를 구명하기 위하여 3T3-L1 전지방세포 분화과정 중 연화 열수 및 에탄올 추출물에 의한 지방축적과 ROS 생성을 관찰하였다. 연화 열수 및 에탄올 추출물은 XTT assay에서 100, 200 및 $400{\mu}g/mL$ 농도에서 세포 독성을 보이지 않았다. 지방세포 분화 중 세포 내 지방축적 및 ROS 생성량을 비교한 결과, 연화 열수 및 에탄올 추출물을 처리한 지방세포의 경우 지방축적량과 ROS 생성량 모두 유의적으로 억제되는 것으로 나타났다. 특히 연화 열수 및 에탄올 추출물을 처리함으로써 지방세포분화와 관련된 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$ 및 aP mRNA의 발현을 유의적으로 감소시켰으며, ROS의 생성과 관련이 있는 주요 유전자인 NOX4 및 catalase의 유전자발현 또한 유의적으로 감소하였다. 이 결과를 통해 연화 열수 및 에탄올 추출물이 3T3-L1 지방세포 내 중성지방의 축적 억제효과와 더불어 ROS 생성 억제에 효과적으로 작용함을 확인하였다. 따라서 연화 열수 및 에탄올 추출물은 비만과 같은 대사증후군 관련 질환의 개선을 위한 천연물 기능성 소재로의 활용이 기대된다.

• 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.

• Journal of Ginseng Research
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• 제47권4호
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• pp.524-533
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• 2023

Background: Obesity is a risk factor for aging and many diseases, and the disorder of lipid metabolism makes it prominent. This study aims to investigate the effect of ginsenoside Rg1 on aging, lipid metabolism and stress resistance Methods: Rg1 was administered to Caenorhabditis elegans (C. elegans) cultured in NGM or GNGM. The lifespan, locomotory activity, lipid accumulation, cold and heat stress resistance and related mRNA expression of the worms were examined. Gene knockout mutants were used to clarify the effect on lipid metabolism of Rg1. GFP-binding mutants were used to observe the changes in protein expression Results: We reported that Rg1 reduced lipid accumulation and improved stress resistance in C. elegans. Rg1 significantly reduced the expression of fatty acid synthesis-related genes and lipid metabolism-related genes in C. elegans. However, Rg1 did not affect the fat storage in fat-5/fat-6 double mutant or nhr-49 mutant. Combined with network pharmacology, we clarified the possible pathways and targets of Rg1 in lipid metabolism. In addition, Rg1-treated C. elegans showed a higher expression of anti-oxidative genes and heat shock proteins, which might contribute to stress resistance Conclusion: Rg1 reduced fat accumulation by regulating lipid metabolism via nhr-49 and enhanced stress resistance by its antioxidant effect in C. elegans.

• 한국임상수의학회지
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• 제29권5호
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• pp.372-376
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• 2012

급성신손상은 높은 이환율과 치사율을 나타내는 심각한 질환이며, 허혈-재관류에 의한 신손상은 급성신손상의 중요한 원인이 된다. 본 연구는 미니돼지에서 급성신손상을 진단하는데 임상적으로 유용한 바이오마커를 찾아내기 위해 실시되었다. 세 마리의 미니돼지에서 60분간 신동맥을 결찰하여 양측성 신허혈을 유도하였다. 각 미니돼지에서 결찰 전, 결찰 후 0, 1, 3, 5일에 혈액 및 뇨 검체를 채취하였다. 혈청 및 뇨 검체에서 BUN, creatinine, 나트륨 및 요산을 측정한 후 나트륨 및 요산의 분획배설율($FE_{Na}$, $FE_{UA}$)을 산출하였다. 또한 IL-6, IL-18, L-FABA 및 GST를 Western immunoblotting을 실시하여 측정하였다. 결과에서 세 마리 미니돼지 모두 혈청 BUN과 creatinine 농도는 결찰 후 1일째에 유의적으로 증가하였다. 그러나 $FE_{Na}$와 $FE_{UA}$는 현저한 개체차를 보였다. 수술 전과 후를 비교했을 때 허혈 이후의 뇨 검체에서는 IL-6, IL-18, L-FABP 및 GST의 농도가 유의적으로 증가하였다. 결론적으로, $FE_{Na}$와 $FE_{UA}$에 대해서는 추가적인 연구가 필요하다고 생각되며, 혈청 BUN, creatinine과 뇨 IL-6, IL-18, L-FABP 및 GST는 돼지의 허혈-재관류 모델에서 다른 바이오마커와 함께 급성신손상을 진단하는 민감한 바이오마커가 될 수 있을 것이라 생각된다.