Enhanced 3T3-L1 Differentiation into Adipocytes by Pioglitazone Pharmacological Activation of Peroxisome Proliferator Activated Receptor-Gamma (PPAR-γ)

Teixeira, Catarina; Sousa, André; Santos, Inês; Rocha, Ana Catarina; Alencastre, Inês S.; Pereira, Ana Cláudia; Martins-Mendes, Daniela; Barata, Pedro; Baylina, Pilar; Fernandes, Rita

doi:10.3390/biology11060806

Cited by 8 publications

(7 citation statements)

References 26 publications

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“…The dose-dependent increase of PPARγ and FABP4 mRNA and protein underscores the role of EF in enhancing adipocyte differentiation. The effect of EF on 3T3-L1 preadipocyte differentiation is similar to that of well-established anti-diabetes drugs, such as pioglitazone and rosiglitazone from the thiazolidinediones group, particularly for promoting the expression of PPARγ and adiponectin [ 49 ]. Pioglitazone enhances peripheral insulin sensitivity.…”

Section: Discussionmentioning

confidence: 76%

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Methanol extract of Elsholtzia fruticosa promotes 3T3-L1 preadipocyte differentiation

Shrestha,

Kim,

Shrestha

et al. 2024

J Anim Sci Technol

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Elsholtzia fruticosa (EF) is present in tropical regions throughout South Asian countries as well as the Himalayas. Although it has been used as a traditional medicine to treat digestive, respiratory, and inflammatory issues, its effect on preadipocyte differentiation is unknown. In this study, we examined the effects of a methanol extract prepared from EF on the differentiation of 3T3-L1 preadipocytes. Cell differentiation was assessed by microscopic observation and oil-red O staining. The expression of adipogenic and lipogenic genes, including PPARγ and C/EBPα , was measured by western blot analysis and quantitative real-time polymerase chain reaction (qRT–PCR), to provide insight into adipogenesis and lipogenesis mechanisms. The results indicated that EF promotes the differentiation of 3T3-L1 preadipocytes, with elevated lipid accumulation occurring in a concentration-dependent manner without apparent cytotoxicity. EF enhances the expression of adipogenic and lipogenic genes, including PPARγ , FABP4 , adiponectin , and FAS , at the mRNA and protein levels. The effect of EF was more pronounced during the early and middle stages of 3T3-L1 cell differentiation. Treatment with EF decreased C/EBP homologous protein (CHOP) mRNA and protein levels, while increasing C/EBPα and PPARγ expression. Treatment with EF resulted in the upregulation of cyclin E and CDK2 gene expression within 24 h, followed by a decrease at 48 h, demonstrating the early-stage impact of EF. A concomitant increase in cyclin-D1 levels was observed compared with untreated cells, indicating that EF modulates lipogenic and adipogenic genes through intricate mechanisms involving CHOP and cell cycle pathways. In summary, EF induces the differentiation of 3T3-L1 preadipocytes by increasing the expression of adipogenic and lipogenic genes, possibly through CHOP and cell cycle-dependent mechanisms.

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Section: Discussionmentioning

confidence: 76%

“…Pioglitazone enhances peripheral insulin sensitivity. Pioglitazone up-regulates adiponectin receptor 2 in 3T3-L1 adipocytes, whereas rosiglitazone increases GLUT-4 expression in muscle cells [ 49 ]. Our results also highlight the unique properties of EF, including the inhibition of CHOP expression at the mRNA and protein level ( Figs.…”

Section: Discussionmentioning

confidence: 99%

Methanol extract of Elsholtzia fruticosa promotes 3T3-L1 preadipocyte differentiation

Shrestha,

Kim,

Shrestha

et al. 2024

J Anim Sci Technol

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“…In addition, we explored the possibility that the differentiation of 3T3-L1 cells into adipocytes might be affected by 8-MNA through interfering with PPARγ pathway, a key transcriptional regulator for adipocyte differentiation [ 12 , 21 ] (Supplementary Fig. 1 ).…”

Section: Resultsmentioning

confidence: 99%

Cellular responses to 8-methyl nonanoic acid, a degradation by-product of dihydrocapsaicin, in 3T3-L1 adipocytes

Wichai

Keawsomnuk

Thongin

et al. 2023

BMC Complement Med Ther

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Background Capsaicinoids, such as dihydrocapsaicin (DHC), exert the health-promoting effects of chili peppers on energy metabolism. The metabolic responses to capsaicinoids are primarily mediated through transient receptor potential cation channel subfamily V member 1 (TRPV1). However, the varying contributions of their metabolites to beneficial health outcomes remain unclear. 8-methyl nonanoic acid (8-MNA), a methyl-branched medium chain fatty acid (MCFA), is an in vivo degradation by-product of DHC. Since MCFAs have emerged as metabolic modulators in adipocytes, here we examined various cellular responses to 8-MNA in 3T3-L1 adipocytes. Methods The viability of 3T3-L1 adipocytes exposed to various concentrations of 8-MNA was assessed by the Calcein AM assay. Biochemical assays for lipid accumulation, AMP-activated protein kinase (AMPK) activity, lipolysis and glucose uptake were performed in 3T3-L1 adipocytes treated with 8-MNA during 48-h nutrient starvation or 5-day maturation. Results 8-MNA caused no impact on cell viability. During nutrient starvation, 8-MNA decreased lipid amounts in association with AMPK activation, a molecular event that suppresses lipogenic processes. Moreover, 3T3-L1 adipocytes that were treated with 8-MNA during 5-day maturation exhibited a reduced lipolytic response to isoproterenol and an increased glucose uptake when stimulated with insulin. Conclusions These results suggest that 8-MNA derived from DHC modulates energy metabolism in adipocytes and also support the idea that the metabolic benefits of chili consumption are partly attributable to 8-MNA.

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“…Two methods of differentiation were applied to the 3T3-L1 cell line (mouse pre-adipocytes), following previously published works [17]. Briefly, the 3T3-L1 cell line was subjected to a 15-day differentiation process using three distinct differentiation mediums (Medium of differentiation I-MD I; Medium of differentiation II-MD II; Medium of differentiation III-MD III).…”

Section: Adipocyte Differentiation and Secretome Collectmentioning

confidence: 99%

Exploring the Potential Protective Effect of Probiotics in Obesity-Induced Colorectal Cancer: What Insights Can In Vitro Models Provide?

Viana,

Rocha,

Sousa

et al. 2024

Applied Sciences

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Colorectal cancer (CRC) is the third most common cancer diagnosed today and the third leading cause of death among cancer types. CRC is one of the gastrointestinal tumors with obesity as the main extrinsic risk factor, since, according to authors, the meta-inflammation sustained by the excess adipose tissue can provide abundant circulating lipids, as well as hormones and metabolites crucial to tumor development and aggressiveness. The gut microbiota can protect the colon from meta-inflammation and endocrine changes caused by obesity. The present study aimed to investigate the antitumor activity of a commercial probiotic in intestinal tumor cells under two adiposity conditions. Experimental assays were performed on the Caco2 cell line (colon adenocarcinoma) supplemented with differentiated adipocyte’s secretomes of the 3T3-L1 cell line (mouse pre-adipocytes) in two adiposity conditions: (i) differentiation without the use of Pioglitazone (noPGZ) and (ii) differentiation using Pioglitazone (PGZ). The Caco2 cells were first exposed to both secretomes for 24 h and evaluated and subsequently exposed to probiotic extract followed by secretome and evaluated. The effects of these treatments were evaluated using cytotoxicity assays by MTT, cell migration by injury, and antioxidant activity by glutathione assay. The use of secretomes showed a statistically significant increase in cell viability in Caco2 cells, either in noPGZ (p < 0.01) or PGZ (p < 0.05), and the probiotic was not able to reduce this effect. In the injury assay, secretome increased cell migration by more than 199% in both adiposity conditions (p < 0.001 in noPGZ and p < 0.01 in PGZ). In the probiotic treatment, there was a reduction in cell migration compared to the control in adiposity conditions. The antioxidant response of Caco2 cells was increased in both adiposity conditions previously exposed to the probiotic supernatant. This pilot work brings to light some findings that may answer why the modulation of the intestinal microbiota using probiotics is an alternative strategy leading to improvements in the condition and stage of the colon tumor. Additional studies are needed to clarify the role of Pioglitazone in this type of tumor and the metabolites of obesity that are attenuated by the use of probiotics.

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Enhanced 3T3-L1 Differentiation into Adipocytes by Pioglitazone Pharmacological Activation of Peroxisome Proliferator Activated Receptor-Gamma (PPAR-γ)

Cited by 8 publications

References 26 publications

Methanol extract of Elsholtzia fruticosa promotes 3T3-L1 preadipocyte differentiation

Methanol extract of Elsholtzia fruticosa promotes 3T3-L1 preadipocyte differentiation

Cellular responses to 8-methyl nonanoic acid, a degradation by-product of dihydrocapsaicin, in 3T3-L1 adipocytes

Exploring the Potential Protective Effect of Probiotics in Obesity-Induced Colorectal Cancer: What Insights Can In Vitro Models Provide?

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