• Herbal Formula Science
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• v.14 no.2
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• pp.86-96
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• 2006

Objectives : This study wad designed to elucidate the short term effect of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 Preadipocyte. Methods : 3T3-L1 preadipocytes obtained from Korean Cell line Bank were cultured in a Dulbecco's modified eagle medium(DMED) culture colution containing 10% fetal bovine serum(FBS) and various concentration of aqueous extract of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 preadipocytes were investigate after treatment for 24 hours b measuring MTT, Oil Red O and latate dehydrogenase activity.. Results : The Rossa rugosae Radix extract inhibited significantly the proliferation of 3T3-L1 preadipocytes and tended to increase latate dehydrogenase activity in the media of differentiated 3T3-L1 preadipocytes & matured 3T3-L1 preadipocytes. the extract also inhibit the lipid accumulation of differentiated and maturaion of 3T3-L1 preadipocytes, suggesting that Rossa rugosae Radix has anti-obesity effect: however further in vivo study is needed to demonstrate its pharmacological effects.

• Journal of Korean Medicine for Obesity Research
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• v.23 no.1
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• pp.1-9
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• 2023

Objectives: Noni fruit juice (NFJ) is liquor extracted from Morinda citrifolia (noni) fruit and has been used as an herbal remedy in many countries. However, the NFJ's anti-adipogenic and anti-inflammatory effects on adipocytes are poorly understood. The purpose of this study was to explore the commercially standardized NFJ effects on lipid accumulation throughout 3T3-L1 preadipocytes differentiation and interleukin-1β (IL-1β)-mediated inducible nitric oxide synthase (iNOS) expression in 3T3-L1 preadipocytes. Methods: Cellular lipid accumulation and triglyceride (TG) content in differentiating 3T3-L1 preadipocytes were assessed subsequently via the Oil Red O staining and AdipoRed assay. MTS assay was used to examine NFJ cytotoxicity in (differentiating) 3T3-L1 preadipocytes. Immunoblotting and reverse transcriptase polymerase chain reaction analysis were used to measure the expression levels of target protein and mRNA in (differentiating) 3T3-L1 preadipocytes, respectively. Results: NFJ treatment at 150 μL/mL led to a substantial reduction of fat accumulation and TG content during 3T3-L1 adipogenesis with no discernable impact on the cell viability. Of note, while NFJ treatment (150 μL/mL) largely inhibited the CCAAT/enhancer-binding protein-α (C/EBP-α) and peroxisome proliferator-activated receptor-β (PPAR-β) protein expressions, it did not influence PPAR-γ in differentiating 3T3-L1 preadipocytes. Of interest, treatment with IL-1β at 20 ng/mL for 4 hours elicited in firm induction of iNOS mRNA expression in 3T3-L1 preadipocytes. However, NFJ treatment at 100 or 200 μL/mL greatly attenuated the IL-1β-induced iNOS mRNA expression in 3T3-L1 preadipocytes. Conclusions: NFJ has anti-adipogenic and anti-inflammatory effects on (differentiating) 3T3-L1 preadipocytes which are in part intervened via control of the expression of C/EBP-α, PPAR-β, and iNOS.

• Journal of Korean Medicine for Obesity Research
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• v.22 no.1
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• pp.38-46
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• 2022

Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN's lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 𝜇l/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-𝛽 and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-𝛼, peroxisome proliferator-activated receptor-𝛽/𝛾, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 𝜇l/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-𝛽 and FAS.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.86-92
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• 2007

Hibiscus sabdariffa L., a tropical beverage material, is used commonly as in folk medicine against hypertension, pyrexia, inflammation, liver disorders, and obesity. However, the mechanism by which Hibiscus sabdariffa L. modulates adipogenic differentiation is remained to be elusive. This report was designed to investigate the inhibitory effect of Hibiscus extract on insulin signaling pathway during adipocyte differentiation in 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were differentiated with isobutylmethylxanthine, dexamethasone, and insulin (MDI) and followed by the addition of Hibiscus extract. Treatment with Hibiscus resulted in a decrease of lipid droplet accumulation, which was suppressed by PI-3 kinase inhibitor wortmannin in 3T3-L1 preadipocytes. Also, Hibiscus extract markedly attenuated the mRNA expression of adipogenic transcriptional factor PPAR${\gamma}$ and adipogenic hormon Leptin during adipogenesis. However, it did not affect the expression of adiponectin in 3T3-L1 preadipocytes differentiated with MDI mixture. Furthermore, Adipogenic differentiation by MDI mixture increased the phosphorylation and expression of PI3-Kinase and Akt in 3T3 preadipocytes, which was markedly suppressed by Hibiscus extract treatment. Taken together, our results suggest that Hibiscus extract suppressed the adipogenic differentiation of 3T3 preadipocytes through activation of PI3-Kinase and Akt signaling pathway.

• Journal of Korean Medicine for Obesity Research
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• v.23 no.2
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• pp.69-77
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• 2023

Objectives: Atriplex gmelinii C. A. Meyer is a halophyte belonging to the Chenopodiaceae family, and its young leaves and stems are used as fodder for livestock. The aim of the present study was to investigate the effects of A. gmelinii extract and its solvent fractions on lipid accumulation during adipogenesis of 3T3-L1 preadipocytes. Methods: The samples of A. gmelinii were separately extracted using methylene chloride and methanol. Subsequently, they were combined to formulate the initial extract, which was then partitioned based on polarity to prepare solvent fractions. Oil Red O staining was employed to measure lipid accumulation during the differentiation of 3T3-L1 preadipocytes. To verify cytotoxicity in 3T3-L1 cells, MTT assays were conducted. The expression levels of transcription factors in 3T3-L1 preadipocytes were measured through Western blotting analysis. Results: At 50 ㎍/mL, treatment of A. gmelinii extract and its solvent fractions during the differentiation of 3T3-L1 preadipocytes significantly diminished lipid accumulation with no noteworthy cytotoxicity on cell viability. Additionally, when investigating the biochemical pathways that underlie the prevention of lipid accumulation using solvent fractions, it was found that the n-BuOH and n-hexane fractions significantly decreased the expression of key transcription factors involved in the generation of fat, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP1c). Conclusions: These findings indicate that A. gmelinii can effectively reduce the accumulation of fat in 3T3-L1 adipocytes, making it a potentially valuable material for mitigating and preventing obesity.

• Journal of Life Science
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• v.27 no.10
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• pp.1104-1110
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• 2017

SET1A is a histone H3K4 methyltransferase that catalyzes di- and trimethylation of histone H3 at lysine 4 (H3K4). Mono-, di-, and trimethylations on H3K4 (H3K4me1, H3K4me2, and H3K4me3, respectively) are generally correlated with gene activation. Although H3K4 methylation is associated with the stimulation of adipogenesis of 3T3-L1 preadipocytes, it remains unknown whether SET1A plays a role in the regulation of adipogenesis of 3T3-L1 preadipocytes. Here, we investigated whether SET1A regulates 3T3-L1 preadipocytes' adipogenesis and characterized the mechanism involved in this regulation. SET1A expression increased during 3T3-L1 preadipocytes' adipogenesis. Consistent with the increased SET1A expression, the global H3K4me3 level had also increased on day 2 after the induction of adipogenesis in 3T3-L1 adipocytes. SET1A knockdown using siRNA in 3T3-L1 preadipocytes inhibited 3T3-L1 preadipocytes' adipogenesis, as assessed by Oil Red O staining and the expression of adipogenic genes, indicating that SET1A stimulates the adipogenesis of 3T3-L1 preadipocytes. SET1A knockdown inhibited the cell proliferation of 3T3-L1 cells during mitotic clonal expansion (MCE) via down-regulation of the cell cycle gene cyclin E1, as well as the DNA synthesis gene, dihydrofolate reductase. Furthermore, SET1A knockdown repressed peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) expression during the late stage of adipogenesis. These results indicate that SET1A stimulates MCE and $PPAR{\gamma}$ expression, which leads to the promotion of 3T3-L1 preadipocytes' adipogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1409-1415
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• 2009

Obesity is especially a serious health problem in industrialized countries, because it is considered to be a risk factor associated with the genesis or development of various metabolic diseases, including cardiovascular disease and type 2 diabetes mellitus. The purpose of this study was to investigate the effects of tanshinone IIA from Salvia miltiorrhiza Bunge on induction of apoptossis and inhibition of adipogenesis in in 3T3-L1 preadipocytes and adipocytes. The results demonstrated that tanshinone IIA decreased cell population growth of 3T3-L1 preadipocytes, assessed with the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and LDH (lactate dehydrogenase) assay. Flow cytometric analysis of 3T3-L1 preadipocytes exposed to tanshinone IIA showed that apoptotic cells increased in a timeand dose-dependent manner. Treatment with tanshinone IIA decreased the number of normal cells and increased the number of apoptotic cells in a dose-dependent manner. The induction of apoptosis in 3T3-L1 preadipocytes by tanshinone IIA was mediated through the activation of caspase-3 and Bax, and then through the cleavage of PARP and the down-regulation of Bcl-2. Moreover, tanshinone IIA significantly decreased the amount of intracellular triglycerides and GPDH (glycerol-3-phosphate dehydrogenase) activity in 3T3-L1 adipocytes. Our results suggest that tanshinone IIA efficiently induces apoptosis and inhibits adipogenesis in 3T3-L1 preadipocytes and adipocytes.

• Herbal Formula Science
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• v.14 no.2
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• pp.76-85
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• 2006

Objectives : We Investigated the effect of Dioscoreae Rhizoma(山藥) on the change of the corticosterone and the rectal temperature(直腸溫渡) of the mice induced by starvation stress(創戰 스트레스). Methods : After administration of Dioscoreae Rhizoma (0.25g/kg, 0.5g, 1.0g/kg, 3g/kg) three times, mice were starved. The corticosterone and rectal temperature were measured after 36.5 hours starvation stress. Results : The plasma cortiosterone levels in the S-2, S-3 and S-4 group were decreased significantly comparing with the control group (P<0.01) after 36.5 hours starvation stress. and rectal temperature was decreased in the control goup comparing with the normal group, but there is no significant change in the Dioscoreae Rhizoma treated group. Conclusion : it might be recognized that Dioscoreae Rhizoma has preventive-effect against starvation stress and also it might be needed further study in various viewpoints. Objectives : This study was disegned to elucidate the short term effect of Rossa rugosae Radix on proliferation. differentiation and maturation of 3T3-L1 Preadipocyte. Methods: 3T3-L1 preadipocytes obtained from Korean Cell Line Bank were cultured in a D ulbecco’ s modified eagle medium(MEM) culture solution containing 10% fetal bovine serum(FBS) and various concentrations of aqueous extract of Rossa rugosae Radix.. The short term effect of the extract of Rossa rugosae Radix on proliferation. differentiation and maturation of 3T3-L1 preadipocytes were investigate after treatment for 24 hours by measuring MTT. Oil Red 0 and latate dehydrogenase activity.. Results: The Rossa rugosae Radix extract inhibited significantly the proliferation of 3T3-L1 preadipocytes and tended to increase latate dehydrogenase activity in the media of differentiated 3T3-L1 preadipocytes & matured 3T3-L1 preadipocytes. the extract also inhibit the lipid accumulation of differentiated 3T3-L1 preadipocytes & matuered 3T3-L1 preadipocytes. Conclusions: These results demonstrated that the Rossa rugosae Radjx extract inhibited the proliferation. differentiation and maturation of 3T3-L1 preadipocytes. suggesting that Rossa rugosae Radix has anti-obesity effect: however further in vivo study is needed to demonstrate its pharmacological effects.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1338-1344
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• 2016

The present study was carried out to evaluate the anti-obesity effect of different concentrations of extracts of hot air-dried ramie leaf (HR) and freeze-dried ramie leaf (FR) in 3T3-L1 cells and pig preadipocytes. To analyze the effect on cell proliferation, cells were treated with $25{\mu}g/mL$ or $100{\mu}g/mL$ HR or FR extract for 2 days. Cell differentiation was evaluated by measuring glycerol-3-phosphate dehydrogenase and lipoprotein lipase (LPL) activities and intracellular triglyceride content. Treatment with either HR or FR extracts inhibited the proliferation of 3T3-L1 cells and pig preadipocytes in a dose-dependent manner. HR extract treatment inhibited the differentiation of both cell types more effectively than FR treatment. The extent of triglyceride accumulation decreased significantly in both cells following either HR or FR treatment. Furthermore, LPL activity significantly decreased after treatment with HR or FR extract. These results indicated that HR and FR extracts may inhibit proliferation and differentiation of 3T3-L1 cells and pig preadipocytes. Further studies are needed to explore the anti-obesity effect of HR and FR extracts.

• Journal of Animal Science and Technology
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• v.55 no.1
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• pp.19-23
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• 2013

The current study was carried out to determine the effects of Kohlrabi (Brassica oleracea var. gongylodes) on proliferation and differentiation of pig preadipocytes and $_3T_3-L_1$ cells. Pig preadipocytes were isolated from the backfat of the new-born pigs. Twenty-four hours after seeding, the cells were washed with DMEM/F-12 (designated day 0). To measure the cell proliferation, the cells were treated with 25 ng/ml and 100 ng/ml ethanol extracts of Kohlrabi (peel and flesh) for two days (day 0 ~ 2). To measure differentiation, the cells were treated with Kohlrabi for two days (day 0 ~ 2) and cell differentiation was measured on day 6. Twenty-five ng/ml and 100 ng/ml of Kohlrabi peel decreased proliferation of pig preadipocytes by 4.59% and 17.7%, respectively, compared with the control and Kohlrabi flesh by 11.4% and 19.2%, respectively. However, Kohlrabi did not inhibit cell differentiation. To measure the effects of Kohlrabi on proliferation and differentiation of $_3T_3-L_1$ cells, the cells were treated with Kohlrabi for two days in culture, like pig preadipocytes. Kohlrabi (both peel and flesh) did not show any effects on cell proliferation and differentiation. In summary, the results of the current study showed that Kohlrabi decreased proliferation of pig preadipocytes, but no inhibitory effects on differentiation of the cells. Kohlrabi had no effects on proliferation and differentiation of $_3T_3-L_1$ cells.