• 한방비만학회지
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• 제23권2호
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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.

• Biomolecules & Therapeutics
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• 제30권5호
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• pp.418-426
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• 2022

Chimeric antigen receptor T (CAR-T) cell therapy is one of the promising anticancer treatments. It shows a high overall response rate with complete response to blood cancer. However, there is a limitation to solid tumor treatment. Additionally, this currently approved therapy exhibits side effects such as cytokine release syndrome and neurotoxicity. Alternatively, bispecific antibody is an innovative therapeutic tool that simultaneously engages specific immune cells to disease-related target cells. Since programmed death ligand 1 (PD-L1) is an immune checkpoint molecule highly expressed in some cancer cells, in the current study, we generated αCD3xαPD-L1 bispecific antibody (BiTE) which can engage T cells to PD-L1⁺ cancer cells. We observed that the BiTE-bound OT-1 T cells effectively killed cancer cells in vitro and in vivo. They substantially increased the recruitment of effector memory CD8⁺ T cells having CD8⁺CD44⁺CD62L^low phenotype in tumor. Interestingly, we also observed that BiTE-bound polyclonal T cells showed highly efficacious tumor killing activity in vivo in comparison with the direct intravenous treatment of bispecific antibody, suggesting that PD-L1-directed migration and engagement of activated T cells might increase cancer cell killing. Additionally, BiTE-bound CAR-T cells which targets human Her-2/neu exhibited enhanced killing effect on Her-2-expressing cancer cells in vivo, suggesting that this could be a novel therapeutic regimen. Collectively, our results suggested that engaging activated T cells with cancer cells using αCD3xαPD-L1 BiTE could be an innovative next generation anticancer therapy which exerts simultaneous inhibitory functions on PD-L1 as well as increasing the infiltration of activated T cells having effector memory phenotype in tumor site.

• Journal of Animal Science and Technology
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• 제62권6호
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• pp.854-863
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• 2020

This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

• 한방비만학회지
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• 제23권1호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제29권3호
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• pp.419-427
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• 2016

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

• Journal of Applied Biological Chemistry
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• 제63권2호
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• pp.169-174
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• 2020

The objective of this study was to investigate the effects of a hypodermic injectable solution comprised of an LPM LB meso solution containing Melissa officinalis L. leaf extract (LPM) on the lipogenesis in the 3T3-L1 cells line. The lipid accumulation measured by oil red o staining in the 3T3-L1 adipocytes treated with LPM, which was reduced in a dose dependent manner and showed 91.7 to 62.9% compared to control group. Its effectiveness with a 50% solution was significantly higher than the hydroxycitric acid (positive control) treatment without showing cell cytotoxicity. In a quantitative real-time PCR, it was demonstrated that the LPM treatment appeared to upregulate the mRNA expression of the adipogenesis-related genes, which included the peroxisome proliferator-activated receptor gamma (50% concentration) while down-regulating the CCAAT-enhancer binding protein alpha (50% concentration) and the sterol regulatory element-binding protein 1c (10, 25, and 50% concentrations). The results from the current study suggest that the LPM could be useful biomaterials that can inhibit obesity in the 3T3-L1 cells, which could possibly be by regulating the specific adipogenic gene transcription factors.

• 동의생리병리학회지
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• 제23권6호
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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.

• Nutrition Research and Practice
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• 제5권3호
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• pp.192-197
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• 2011

The dietary intake of whole grains is known to reduce the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. To investigate whether there are anti-adipogenic activities in various Korean cereals, we assessed water extracts of nine cereals. The results showed that treatment of 3T3-L1 adipocytes with Sorghum bicolor L. Moench, Setaria italica Beauvois, or Panicum miliaceum L. extract significantly inhibited adipocyte differentiation, as determined by measuring oil red-O staining, triglyceride accumulation, and glycerol 3-phosphate dehydrogenase activity. Among the nine cereals, P. miliaceum L. showed the highest anti-adipogenic activity. The effects of P. miliaceum L. on mRNA expression of peroxisome proliferator-activated receptor-${\gamma}$, sterol regulatory element-binding protein 1, and the CCAAT/enhancer binding protein-${\alpha}$ were evaluated revealing that the extract significantly decreased the expression of these genes in a dose-dependent manner. Moreover, P. miliaceum L. extract changed the ratio of monounsaturated fatty acids to saturated fatty acids in adipocytes, which is related to biological activity and cell characteristics. These results suggest that some cereals efficiently suppress adipogenesis in 3T3-L1 adipocytes. In particular, the effect of P. miliaceum L. on adipocyte differentiation is associated with the downregulation of adipogenic genes and fatty acid accumulation in adipocytes.

• Journal of Nutrition and Health
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• 제47권1호
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• pp.1-11
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• 2014

널리 음용되고 있는 녹차의 EGCG과 우리나라 국민의 상당수가 복용하고 있는 건강기능성 식품 성분인 글루코사민은 이전의 연구들을 통해서 지방세포의 분화를 억제하는데 효과가 있다고 보고되어왔다. 이 두 물질의 병합처리로 기대되어지는 지방세포에서의 adipogenesis 및 지방축적감소에 대한 상승효과는 검증된 바 없으며, 효과에 대한 cell cycle 차원에서의 접근은 없었다. 본 연구 결과에서 EGCG와 Glucosamine 6-phosphate는 adipogenesis 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1에 대한 직접적인 발현 억제 뿐아니라, $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1와 매개된 FAS, ACSL1, LPL과 같은 adipogenic target 유전자의 발현 감소를 통하여 지방세포의 분화와 지방세포 내 지방축적을 감소시키는 효과를 나타냈다. 그리고 HSL과 perilipin의 발현조절을 통해 부분적인 lipolytic effet도 나타냈다. 또한 지방세포의 분화가 개시되는데 있어 중요한 DNA의 remodeling 과정인 mitotic clonal expansion (MCE) 과정 중 G0/G1 phase 단계에서 cell cycle 정지 유도와 그로인한 S phase 및 G2/M phase로 세포주기이행의 방해를 통해 지방세포가 분화되는 것은 억제하였다. 이러한 효과들은 EGCG 농도가 높아질수록, 그리고 EGCG를 단독으로 처리한경우보다 Glucosamine 6-phosphate와 병합하였을 때 효과적이었다. 따라서 EGCG 단독처리 및 glucosamine 6-phosphate와의 병합처리는 지방세포에서 adipogenesis와 adipogenic관련 유전자들의 발현 억제 및 MCE 단계의 cell cycle arrest를 통해 지방세포의 분화를 억제하고 지방축적을 감소시켜 항비만 효과를 나타냈으며, 이러한 효과는 두 성분의 병합처리에서 조금 더 효과적이었다고 할 수 있다. 비록 두 성분의 병합처리가 기대했던 만큼은 아니었으나 항비만 효과에 대한 상승효과가 있다고 볼 수 있다.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1895-1902
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• 2008

Adipose tissue is an important endocrine organ involved in the control of whole body energy homeostasis and insulin sensitivity. Considering the increased incidence of obesity and obesity-related disorders, including diabetes, it is important to understand thoroughly the process of adipocyte differentiation and its control. Therefore, we performed a differential proteome mapping strategy using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to identify intracellular proteins that are differentially expressed during adipose conversion of 3T3-L1 pre-adipocytes in response to an adipogenic cocktail. In the current study, we identified 46 differentially expressed proteins, 6 of which have not been addressed previously in 3T3-L1 cell differentiation. Notably, we found that phosphoribosyl pyrophosphate synthetase (PRPS), a regulator of cell proliferation, was preferentially expressed in pre-adipocytes than in fully differentiated adipocytes. In conclusion, our results provide valuable information for further understanding of the adipogenic process.