Potential miRNA Involvement in the Anti-Adipogenic Effect of Resveratrol and Its Metabolites

Abstract

Objective

Scientific research is constantly striving to find molecules which are effective against excessive body fat and its associated complications. Taking into account the beneficial effects that resveratrol exerts on other pathologies through miRNA, the aim of the present work was to analyze the possible involvement of miRNAs in the regulation of adipogenic transcription factors peroxisome proliferator-activated receptor y (ppar gamma), CCAAT enhancer-binding proteins a and beta (cebp beta and cebpa) induced by resveratrol and its metabolites.

Methods

3T3-L1 maturing pre-adipocytes were treated during differentiation with 25 mu M of trans-resveratrol (RSV), trans-resveratrol-3-O-sulfate (3S), trans-resveratrol-3'-O-glucuronide (3G) and trans-resveratrol-4'-O-glucuronide (4G). After computational prediction and bibliographic search of miRNAs targeting ppar gamma, cebp beta and cebpa, the expression of microRNA-130b-3p (miR-130b-3p), microRNA-155-5p (miR-155-5p), microRNA-27b-3p (miR-27b-3p), microRNA-31-5p (miR-31-5p), microRNA-326-3p (miR-326-3p), microRNA-27a-3p (miR27a-3p), microRNA-144-3p (miR-144-3p), microRNA-205-5p (miR-205-5p) and microRNA224-3p (miR-224-3p) was analyzed. Moreover, other adipogenic mediators such as sterol regulatory element binding transcription factor 1 (srebfl), kriippel-like factor 5 (k1f5), liver x receptor a (lxra) and cAMP responding element binding protein 1 (crebl), were measured by Real Time RT-PCR. As a confirmatory assay, cells treated with RSV were transfected with anti-miR-155 in order to measure cebp beta gene and protein expressions.

Results

Of the miRNAs analyzed only miR-155 was modified after resveratrol and glucuronide metabolite treatment. In transfected cells with anti-miR-155, RSV did not reduce cebp beta gene and protein expression. 3S decreased gene expression of crebl, klf5, srebfl and lxra.

Conclusions

While RSV and glucuronide metabolites exert their inhibitory effect on adipogenesis through miR-155 up-regulation, the anti-adipogenic effect of 3S is not mediated via miRNAs.