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Inhibition of adipogenesis by RGD-dependent disintegrin
Yu-Ting Lin1, Chih-Hsin Tang, Woei-Jer Chuang
1Department of Pharmacology, College of Medicine, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei, Taiwan.
Abstract:
Adipogenesis plays a central role in obesity development. The processes of adipogenesis include migration, adhesion, proliferation and survival of preadipocytes and differentiation to mature adipocytes. Many of these biological functions are related to integrins. Here, we found that snake venom-derived arginine-glycine-aspartic acid (RGD)-containing disintegrin inhibited adipogenesis. Rhodostomin but not rhodostomin RGD mutants (RGE-Rn and AKGDWN-Rn) caused the detachment of primary cultured preadipocyte. Furthermore, rhodostomin also inhibited focal adhesion of preadipocyte, including the inhibition of the expression of focal adhesion kinase (FAK) and FAK phosphorylation, assembly of vinculin and reorganization of actin cytoskeleton. Cell viability of preadipocytes was decreased after rhodostomin treatment in a concentration-dependent manner. The results of flow cytometric analysis showed that rhodostomin induced cell apoptosis. In addition, chromatin condensation was observed in DAPI staining. The increase of Bax expression and activation of capsase-3 was detected following rhodostomin treatment. Addition of dexamethasone, IBMX and insulin induced differentiation of preadipocytes into mature adipocytes and treatment of cells with rhodostomin during the initial 3 days showed less mature adipocytes following 9-10 days of differentiating period. The triglyceride content and gene expression of peroxisome proliferators-activated receptor gamma (PPARgamma) and leptin also decreased in response to the treatment of rhodostomin. These results suggest that disintegrin inhibits processes of adipogenesis and may be developed to treat obesity.
Insights
Snake venom disintegrins, like rhodostomin, inhibit adipogenesis by disrupting preadipocyte adhesion and promoting apoptosis. This suggests potential therapeutic applications for obesity treatment.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Medicine
Background:
- Adipogenesis, the process of fat cell formation, is crucial in obesity development.
- Integrins play a key role in preadipocyte functions like migration, adhesion, and survival.
- Snake venom-derived disintegrins containing the RGD motif are known to interact with integrins.
Purpose of the Study:
- To investigate the effect of snake venom-derived disintegrins on adipogenesis.
- To determine the specific mechanisms by which rhodostomin inhibits adipogenesis.
- To evaluate the potential of rhodostomin as an anti-obesity therapeutic.
Main Methods:
- Primary cultured preadipocytes were treated with rhodostomin and its mutants.
- Assays included cell detachment, focal adhesion analysis (FAK, vinculin, actin cytoskeleton), cell viability, apoptosis markers (flow cytometry, DAPI, Bax, caspase-3), and adipogenic differentiation.
- Gene expression analysis of PPARgamma and leptin, and triglyceride content were measured.
Main Results:
- Rhodostomin, but not its RGD mutants, caused preadipocyte detachment and inhibited focal adhesion.
- Rhodostomin decreased preadipocyte viability, induced apoptosis, and inhibited adipogenic differentiation.
- Treatment with rhodostomin reduced triglyceride content and suppressed PPARgamma and leptin expression.
Conclusions:
- Snake venom-derived disintegrins, specifically rhodostomin, effectively inhibit adipogenesis.
- Rhodostomin disrupts key cellular processes involved in adipogenesis, including adhesion and survival.
- Rhodostomin shows promise as a potential therapeutic agent for treating obesity.
