• Animal Bioscience
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• v.35 no.2
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• pp.196-203
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• 2022

Objective: This study compared differentially expressed genes (DEGs) between groups with high and low numbers of fine marbling particles (NFMP) in the longissimus thoracis muscle (LT) of Korean cattle to understand the molecular events associated with fine marbling particle formation. Methods: The size and distribution of marbling particles in the LT were assessed with a computer image analysis method. Based on the NFMP, 10 LT samples were selected and assigned to either high- (n = 5) or low- (n = 5) NFMP groups. Using RNA sequencing, LT transcriptomic profiles were compared between the high- and low-NFMP groups. DEGs were selected at p<0.05 and |fold change| >2 and subjected to functional annotation. Results: In total, 328 DEGs were identified, with 207 up-regulated and 121 down-regulated genes in the high-NFMP group. Pathway analysis of these DEGs revealed five significant (p<0.05) Kyoto encyclopedia of genes and genomes pathways; the significant terms included endocytosis (p = 0.023), protein processing in endoplasmic reticulum (p = 0.019), and adipocytokine signaling pathway (p = 0.024), which are thought to regulate adipocyte hypertrophy and hyperplasia. The expression of sirtuin4 (p<0.001) and insulin receptor substrate 2 (p = 0.043), which are associated with glucose uptake and adipocyte differentiation, was higher in the high-NFMP group than in the low-NFMP group. Conclusion: Transcriptome differences between the high- and low-NFMP groups suggest that pathways regulating adipocyte hyperplasia and hypertrophy are involved in the marbling fineness of the LT.

• Nutritional Sciences
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• v.6 no.3
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• pp.148-154
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• 2003

Although various raw plant materials have been demonstrated to exert anti-obesity effects to a greater or lesser extent in both humans and animals when they are used to supplement the diet, it has not been shown extensively that they influence adipocyte cell differentiation involving lipid metabolic gene expressions. Using a well-established 3T3-L1 preadipocyte differentiation system, we decided to look into molecular and cellular event occurring during adipocyte differentiation when raw plant materials aye included in the process, in an effort to demonstrate the potential use of a screening system to define the functions of traditionally well-known materials. To these ends, the effects of ethanol (EtOH) or EtOH/distilled water (DW) extracts of Wax Gourd were examined using cytochemical and molecular analyses to determine whether components of the extracts modulate adipocyte differentiation of 3T3-Ll preadipocytes in vitro. The cytochemical results demonstrated that EtOH or EtOH/DW extracts did not affect lipid accumulation and cell proliferation, although the degree of lipid accumulation was influenced slightly depending on the extract. EtOH extract was highly effective in apoptotic induction during differentiation of 3T3-Ll preadipocytes (p<0.05). Reverse transcription-polymerase chain reaction (RT-PCR) analysis of lipoprotein lipase (LPL), Uncoupling protein (Ucp) 2, 3 and 4 also showed that while LPL expression was not influenced, Ucp2, 3 and 4 were up regulated in the EtOH extract-treated group and down regulated in the EtOH/DW extract-treated group. These changes in gene expressions suggest that the components in different fractions of Wax Gourd extracts may modulate lipid metabolism by either direct or indirect action. Taking these results together, it was concluded that molecular and cellular analyses of adipocyte differentiation involving lipid metabolic genes should facilitate understanding of cellular events occurring during adipocyte differentiation. Furthermore, the experimental scheme and analytical methods used in this study should provide a screening system for the functional study of raw plant materials in obesity research.

• Journal of agriculture & life science
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• v.45 no.2
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• pp.85-92
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• 2011

The current study was conducted to investigate effects of sera taken from Hanwoo at different age on adipocyte differentiation. Sera were taken from Korean native (Hanwoo) steers at 6, 12, 18, and 24 months of age, respectively and supplemented to 3T3-L1 preadipocytes after the cells reached confluence and maintained 10 days thereafter. For the first 2 days (48 h), cells were induced to differentiate by addition of differentiation factors, methylisobutylxanthine, daxamethasone, and insulin. After the differentiation, the cells were incubated without differentiation factors except insulin. The cells lost their fibroblastic shape and showed round-up appearances after 10 days incubation with FBS and the sera of Hanwoo steers. Big lipid droplets appeared in the cells cultured with FBS and the sera taken from Hanwoo at 18 and 24 months of age. After 18 months of age, GPDH activity was statistically higher than 6 and/or 12 months of age (P < 0.05). Based on morphology and Sn-glycerol-3-phosphate dehydrogenase activities, Hanwoo steers expressed aged-dependent adipogenic activities, indicating that aged sera may result in high adipocyte differentiation. It is concluded that the 18 months of age may be 'threshold' to express major adipogenic activities. This may strongly support previous field studies reporting considerable increase in fat contents of Hanwoo carcass at over 18 months of age.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.651-663
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• 2016

Corn silk, Job's tears, Lentinus edodes, and apple peel 70% ethanol extracts (CS, JT, LE, and AP) were studied for their antioxidant activities. CS among all extracts showed the highest antioxidant activities based on total polyphenol and flavonoid contents, 2,2-diphenyl-${\beta}$-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical scavenging activity, and reducing power. Adipocyte differentiation was investigated by Oil Red O staining assay using CS, JT, LE, AP, and extract of developed bread containing corn silk, Job's tears, Lentinus edodes, and apple peel (DB) treated to 3T3-L1 adipocytes. DB1 and DB2 showed anti-adipogenic and antioxidant effects. Triglyceride (TG) accumulation in 3T3-L1 cells was measured, and among the samples tested (CS, JT, LE, and AP), CS was found to have the highest inhibitory activity against TG accumulation of differentiated 3T3-L1 adipocytes and regulated factors associated with adipogenesis. CS suppressed lipid droplet formation and adipocyte differentiation in 3T3-L1 cells in a dose-dependent manner. We examined the effects of CS on the levels of CCAAT-enhancer-binding protein ${\beta}(C/EBP{\beta})$, peroxisome proliferator activated receptor ${\gamma}(PPAR{\gamma})$, and adipocyte-specific lipid binding protein (aP2) mRNA as well as protein levels in 3T3-L1 cells treated with CS at various concentrations (0, 10, 50, and $100{\mu}g/mL$) during adipocyte differentiation and treatment with CS in 3T3-L1 adipocytes down-regulated expression of $PPAR{\gamma}$ and aP2 mRNA. CS also significantly inhibited up-regulation of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 proteins during adipocyte differentiation. These data indicate that DBs have anti-adipogenic activity induced by CS in 3T3-L1 preadipocytes, and CS exerts anti-adipogenic activity by inhibiting expression of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 signaling pathway in 3T3-L1 adipocytes. JT, LE, and AP had no inhibitory effects on differentiation of 3T3-L1 preadipocytes but displayed strong antioxidant effects. These results suggest that the developed bread may be a health beneficial food that can prevent or treat obesity and diseases induced by oxidative stress.

• The Journal of Korean Medicine
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• v.26 no.2 s.62
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• pp.241-251
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• 2005

Objectives: This experimental study was designed to investig:ue the effects of Oryungsan-gagampang on leptin and leptin receptor levels and differentiation of 3T3-L1 adipocyte. Methods: After 3T3-L1 adipocytes were incubated with various concentrations of Oryungsan-gagampang and Reductil(r) for 7 days, leptin and leptin receptor levels in 3T3-Ll adipocytes were measured by ELISA. To elucidate the mechanism of inhibitory effects of Oryungsan-gagampang on obesity, the 3T3-L1 adipocytes after oil red 0 staining were taken by digital photo system. Results: 1. Oryungsan-gagampang $1,000{\mu}g/ml$ significantly increased leptin levels in 3T3-L1 adipocytes in comparison with the control group (p<0.05), and Oryungsan-gagampang 0.1 10, $1,000{\mu}g/ml$ significantly increased leptin receptor levels in 3T3-L1 adipocytes in comparison with the control group (p<0.05). 2. Oryungsan-gagampang inhibited of differentiation of 3T3-L1 adipocytes. Conclusions: Oryungsan-gagampang showed significant effects on inhibiting differentiation of 3T3-Ll adipocytes, and increasing leptin levels and leptin receptor levels in 3T3-L1 adipocytes. Therefore, Oryungsan-gagampang could be used to treat obesity, but further studies are required.

• The Korea Journal of Herbology
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• v.31 no.4
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• pp.11-18
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• 2016

Objectives : The objective of this study was to investigate the effect of Myrrh 80% ethanol extract on adipocyte differentiation and adipogenesis in 3T3-L1 cell.Methods : Myrrh was prepared by extracting with 80% ethanol. Cell viability was assessed by MTT assay using 3T3-L1 cells. Anti-obesity activity was measured in lipid droplets and triglyceride (TG) accumulation in 3T3-L1 cells. We also analyzed the expression of C/EBPβ, C/EBPα, PPARγ, SREBP1c, and aP2 by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, we observed the production of fatty acid, acetyl-CoA carboxylase and Oil-red O stainingResults : No cytotoxicity from Myrrh 80% ethanol extracts was observed at the concentration of 1, 10, 100 (㎍/㎖) in 3T3-L1 cells. Treatment with Myrrh significantly suppressed the terminal differentiation of 3T3-L1 in a dose-dependent manner, as confirmed by a decrease in triglyceride and Fatty acid and Acetyl-CoA carboxylase. Also, Myrrh exhibited potential adipogenesis inhibition and downregulated the expression of pro-adipogenic transcription factors, such as sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα) and C/EBPβ, and adipocyte expressed genes, such as adipocyte fatty acid binding protein (aP2) and Fas. In addition, lipid accumulation determined by Oil-red O staining showed that Myrrh extract had inhibitory effects on lipid accumulation in 3T3-L1 cells.Conclusions : These results suggest that Myrrh suppresses obesity factors in 3T3-L1 cells. Myrrh may be a useful medical herbs for attenuating metabolic diseases such as obesity.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.721-726
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• 2009

The molecular mechanism of adipocyte differentiation has been well documented. However, the effect of specific nutrients such as lysine on adipocyte differentiation is poorly understood especially in ruminant animals. Therefore, the aim of the present study was to elucidate the influence of lysine on adipocyte differentiation and adipogenic genes in cultured bovine preadipocytes. The preadipocytes were treated with different concentrations of lysine (40, 160, 320 mg/L) or troglitazone (10 ${\mu}M$) for 2 days and then subsequently cultured in differentiation medium until day 6. Expression levels of $C/EBP{\alpha}$ were significantly higher (p<0.001) in 40 and 160 mg/L lysine-treated cells compared to 320 mg/L treatment. Though there was an increasing trend in $PPAR{\gamma}$ expression levels with the decreasing lysine concentration, the results were not significant. The preadipocyte factor (pref-1), expression significantly (p<0.001) reduced with decreasing lysine concentration. The Oil red O staining results were better in 40 mg/L treated cells compared to 160 and 320 mg/L lysine treated cells. Our overall results indicate that insufficient supply of lysine increases the adipogenic potential in bovine intramuscular preadipocytes.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.928-931
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• 2009

Effects of perilla leaf extracts (PLE) on adipocytes differentiation of 3T3-L1 cells were examined. Ethanol extract of PLE treatment significantly decreased lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. Moreover, gene expression levels of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), the key adipogenic transcription factor, were markedly decreased by PLE. PLE also suppressed adipocyte fatty acid binding protein (aP2) and glycerol-3-phosphate dehydrogenase (GPDH), which are adipogenic marker proteins. These results suggest that PLE treatment suppressed differentiation of 3T3-L1 adipocytes, in part by down-regulating expression of adipogenic transcription factor and other specific target genes.

• Journal of Life Science
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• v.30 no.12
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• pp.1092-1100
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• 2020

The six-transmembrane epithelial antigen of prostate 4 (Steap4) is a metalloreductase that plays a role in intracellular iron and cupper homeostasis, inflammatory response, and glucose and lipid metabolism. Previously, Steap4 has been reported to stimulate adipocyte differentiation; however, the underlying mechanisms of this action remain unexplored. In the present study, we investigated the molecular mechanisms involved in Steap4-induced adipocyte differentiation using 3T3-L1 cells, immortalized brown adipocyte (iBA) cells, and mouse embryonic fibroblast C3H10T1/2 cells. The knockdown of Steap4 using adenovirus-containing shRNA attenuated mitotic clonal expansion (MCE), as evidenced by the impaired proliferation of 3T3-L1 cells, iBA cells, and C3H10T1/2 cells within 48 hr after adding the differentiation medium. Steap4 knockdown downregulated G1/S phase transition-related cell cycle regulators (including cyclin A and cyclin D) and upregulated cell cycle inhibitors (including p21 and p27). Furthermore, Steap4 knockdown inhibited the phosphorylation of p38 mitogen-activated protein kinase, extracellular signal-regulated kinase, and Akt. Moreover, Steap4 knockdown repressed the expression of early adipogenic activators, such as CCAAT-enhancer-binding protein β (C/EBPβ) and Kruppel-like factor family factor 4 (KLF4). On the other hand, Steap4 knockdown stimulated the expression of adipogenic inhibitors, including KLF2, KLF3, and GATA2. The overexpression of Steap4 using an adenovirus removed the repressive histone marks H3K9me2 and H3K9me3 on the promoter of C/EBPβ. These results indicate that Stepa4 stimulates adipocyte differentiation through the induction of MCE and the modulation of early adipogenic transcription factors, including C/EBPβ, during the early phase of adipocyte differentiation.

• Journal of Life Science
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• v.24 no.10
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• pp.1085-1091
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• 2014

The purpose of this study was to determine the effect of selenate on adipocyte differentiation and to identify genes involved in the modulation of adipogenesis in 3T3-L1 cells. To test the effect of selenate on adipocyte differentiation, adipogenesis was induced in cells using various concentrations ($0-100{\mu}M$) of selenate. Various phases of adipogenesis were induced: postconfluent (PC), early phase (EP, d0-d2), postmitotic growth arrest (PM, d2-d4), and all period (AP). The PC cells exposed to selenate for 24 h displayed dose-dependent inhibition of intracellular lipid droplet accumulation on day 6 of adipogenesis. Two days of selenate treatment at EP or AP inhibited adipogenesis, with an approximately 20-80% reduction in lipid accumulation compared to that of a control (p<0.05). When preadipocytes were exposed to selenate during the PM period, the antiadipogenic effect of selenate was attenuated. Two types of selenoprotein genes (Seps1 and Sepp1) were up-regulated by the selenate treatment during mitotic clonal expansion, whereas these genes were down-regulated during PM growth arrest (p<0.05). The findings demonstrate the antiadipogenic function of selenate and the possible involvement of Sepp1 and Seps1 genes in selenate-inhibited adipogenesis in 3T3-L1 cells.