Elastin-derived peptide VGVAPG decreases differentiation of mouse embryo fibroblast (3T3-L1) cells into adipocytes

ABSTRACT

Elastin is a highly elastic protein present in connective tissue. As a result of protease activity, elastin hydrolysis occurs, and during this process, elastin-derived peptides (EDPs) are released. One of the constitutively repeating elastin and EDP building sequences is the hexapeptide VGVAPG. Therefore, the aim of our research was to define the effect of VGVAPG peptide on adipogenesis in a mouse 3T3-L1 cell line. 3T3-L1 cells were differentiated according to a previously described protocol and exposed to increasing concentrations of VGVAPG or VVGPGA peptide. The obtained results showed that VGVAPG peptide does not stimulate reactive oxygen species (ROS) production, caspase-1 activation, and 3T3-L1 cell proliferation. In the second part of the experiments, it was proved that VGVAPG peptide decreased lipid accumulation as measured by oil red O staining, which was confirmed by the profile of increased expression markers of undifferentiated preadipocytes. In our experiments, 10 nM VGVAPG added for differentiating to adipocytes increased the expression of Pref-1, serpin E1, and adiponectin as compared to rosiglitazone (PPARγ agonist)-treated group and simultaneously decreased the expression of VEGF and resistin as compared to the rosiglitazone-treated group. The obtained results show that VGVAPG peptide sustains 3T3 cells in undifferentiated state.

Abbreviations: DMSO: dimethyl sulphoxide; EBP: elastin-binding protein; EDPs: elastin-derived peptides; FBS: foetal bovine serum; Glb1: gene for beta-galactosidase; LDL: low-density-lipoprotein; PAI-1 (Serpin E1): plasminogen activator inhibitor-1; PBS: phosphate-buffered saline; PPARγ: peroxisome proliferator-activated receptor gamma; Pref-1: preadipocyte factor 1; ROS: reactive oxygen species; VEGF-A: vascular endothelial growth factor-A; VGVAPG: Val-Gly-Val-Ala-Pro-Gly; β-Gal: beta-galactosidase; ORO: oil red O; IBMX: 3-isobutyl-1-methylxanthine; H₂DCFDA: 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate; DMEM: Dulbecco’s Modified Eagle’s Medium; VVGPGA: Val-Val-Gly-Pro-Gly-Ala.

KEYWORDS:

• VGVAPG
• elastin-derived peptides
• 3T3
• lipid accumulation

Highlights

• VGVAPG peptide does not stimulate ROS production and activation of caspase-1 in 3T3-L1 cells

• VGVAPG peptide does not induce proliferation of 3T3-L1 cells

• VGVAPG peptide decreases differentiation of 3T3-L1 cells into adipocytes

• VGVAPG peptide decreases lipid accumulation as measured by ORO assay

• VGVAPG peptide increases expression of Pref-1, serpin E1 and adiponectin as compared to rosiglitazone

• VGVAPG peptide decreases expression of VEGF and resistin as compared to rosiglitazone

Acknowledgments

This work was supported by statutory funds from the University of Information Technology and Management in Rzeszow, Poland (DS 503-07-01-21).

Author statement

Konrad A. Szychowski: Conceptualization, Methodology, Validation, Formal analysis, Investigation, Writing - Review & Editing, Project administration, Data Curation, Writing - Original Draft. Bartosz Skóra.: Data curation, Writing- Original draft preparation. Jakub Tobiasz: Investigation. Jan Gmiński: Supervision Writing - Review & Editing, Funding acquisition.

Disclosure statement

The authors declare no conflict of interests.

Additional information

Funding

This work was supported by the University of Information Technology and Management in Rzeszow [DS 503-07-01-21].