Elsevier

Phytomedicine

Volume 21, Issue 5, 15 April 2014, Pages 758-765
Phytomedicine

Trigonelline attenuates the adipocyte differentiation and lipid accumulation in 3T3-L1 cells

https://doi.org/10.1016/j.phymed.2013.11.007Get rights and content

Abstract

Trigonelline is a natural alkaloid mainly found in Trigonella Foenum Graecum (fenugreek) Fabaceae and other edible plants with a variety of medicinal applications. Therefore, we investigated the molecular mechanism of trigonelline (TG) on the inhibition of adipocyte differentiation and lipid accumulation in 3T3-L1 cells. Trigonelline suppressed lipid droplet accumulation in a concentration (75 and 100 μM) dependent manner. Treatment of adipocyte with of TG down regulates the peroxisome proliferator-activated receptor (PPARγ) and CCAAT element binding protein (C/EBP-α) mRNA expression, which leads to further down regulation of other gene such as adiponectin, adipogenin, leptin, resistin and adipocyte fatty acid binding protein (aP2) as compared with respective control cells on 5th and 10th day of differentiation. Further, addition of triognelline along with troglitazone to the adipocyte attenuated the troglitazone effects on PPARγ mediated differentiation and lipid accumulation in 3T3-L1 cells. Trigonelline might compete against troglitazone for its binding to the PPARγ. In addition, adipocyte treated with trigonelline and isoproterenol separately. Isoproterenol, a lipolytic agent which inhibits the fatty acid synthase and GLUT-4 transporter expression via cAMP mediated pathway, we found that similar magnitude response of fatty acid synthase and GLUT-4 transporter expression in trigonelline treated adipocyte. These results suggest that the trigonelline inhibits the adipogenesis by its influences on the expression PPARγ, which leads to subsequent down regulation of PPAR-γ mediated pathway during adipogenesis. Our findings provide key approach to the mechanism underlying the anti-adipogenic activity of trigonelline.

Introduction

Trigonelline (TG) is an alkaloid which is present in Trigonella Foenum Graecum (fenugreek) Fabaceae and other edible plants such as onions, peas, soybeans cantaloupe, corn and coffee which possess the more medicinal properties such as anticarcinogenic (Anthoni et al. 1991), antimigraine, anti-hypercholesterolemia (Abe and Kaneda 1975) and antidiabetic activities (Yoshinari et al. 2009). Trigonelline reduces the diabetic mediated hypertensive nephropathy in rats (Ghule et al. 2012). Trigonelline is a second most abundant alkaloid in green coffee beans. It generates the non-volatile alkylpridiniums during thermal degradations which contributes to the antioxidative activity of roasted coffee (Gomez-Ruiz et al. 2007). TG is a metabolite of niacin in humans, which is a component of vitamin supplements and is used for its antihyperlipidemic and hypocholesterolemic activity (Yuyama and Suzuki 1991).

Obesity is a very serious health problem in worldwide because of obesity is closely associated with the prevalence of diabetic mellitus, severe cardiovascular disease (Hursting and Dunlap 2012) and it induces the metabolic deregulations such as impairment of glucose uptake and fatty liver which ultimately produces hepatic steatosis (Ferre and Foufelle 2010). The reason for obesity is increasing of adipose tissues which lead to fatty acid accumulations in the preexisting adipocytes. In addition to increase the proliferation and differentiations of pre-adipocytes.

Adipogenesis is the process by which pre-adipocyte is converted into differentiated adipocyte (Otto and Lane 2005). During differentiation process involved the many key transcriptional factors such as C/EBP-δ and C/EBP-β join together induce the expression of PPAR-γ and C/EBP-α (White and Stephens 2010). The PPAR-γ and C/EBP-α regulate the gene that involved in the adipogenesis and lipogenesis such as adiponectin (Shehzad et al. 2012), adipogenin (Hong et al. 2005), FAS (Shimano et al. 1996), leptin (Hwang et al. 1997), resistin (Yong and Lazar 2002). The present study is to assess the effect of TG on adipogenic transcriptional and secretory factor gene expressions which are involved in the adipocytes differentiation and lipid accumulation in 3T3-L1 cell line.

Section snippets

Chemicals

Mouse 3T3-L1 pre-adipocytes were purchased from the American Type Culture Collection (ATCC, USA). Dulbecco modified Eagle medium/high glucose, cell viability assay kit (EZ-CYTox, Daeillab Service Co. Ltd.), mRNA extraction kit and qPCR kit was obtained from the Daeil Lab services Co., Ltd. and Invitrogen Life technology. Trigonelline and other analytical grade chemicals were procured from Sigma (USA)

Cell proliferation assay

The water soluble tetrazolium (WST;

Effect of trigonelline on adipocyte proliferation

The adipocytes were treated with different concentrations (25, 50, 75 and 100 μM) of trigonelline (TG) for 24 and 48 h for cell proliferation assay. This result shows that the TG significantly decreased the adipocyte proliferation at a concentration of 100 μM after 24 and 48 h incubation, whereas 75 μM, significantly decreased the cell proliferation after 48 h only (Fig. 1). At the same time 75 and 100 μM of TG did not disturb the normal cell proliferations as compared with control cells. At 24 h

Discussion

Adipocytes differentiations and cytoplasmic lipid accumulations are very closely associated with the development of obesity (Camp et al. 2002). A reduction of obesity is closely related to the inhibition of differentiations and lipid accumulation in the adipocytes (Otto and Lane 2005). The oil red O staining and microscopic results revealed that the adipocytes exposed with trigonelline (TG) remarkably decreased the lipid accumulation in a concentration dependent manner at 5th and 10th of

Acknowledgments

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ007538)” Rural Development Administration, Republic of Korea.

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