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Inhibition of dipeptidyl peptidase 8/9 impairs preadipocyte differentiation.

Han R, Wang X, Bachovchin W, Zukowska Z, Osborn JW - Sci Rep (2015)

Bottom Line: Here we demonstrate that the DPP8/9 selective inhibitor 1G244 blocks adipogenesis in preadipocyte 3T3-L1 and 3T3-F422A, while DPP4 and FAP inhibitors have no effect.In addition, knockdown of DPP8 or DPP9 significantly impairs adipocyte differentiation in preadipocytes.These results indicate the importance of DPP8 and DPP9 on adipogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, USA.

ABSTRACT
Adipocytes are the primary cells in adipose tissue, and adipocyte dysfunction causes lipodystrophy, obesity and diabetes. The dipeptidyl peptidase (DPP) 4 family includes four enzymes, DPP4, DPP8, DPP9 and fibroblast activation protein (FAP). DPP4 family inhibitors have been used for the treatment of type 2 diabetes patients, but their role in adipocyte formation are poorly understood. Here we demonstrate that the DPP8/9 selective inhibitor 1G244 blocks adipogenesis in preadipocyte 3T3-L1 and 3T3-F422A, while DPP4 and FAP inhibitors have no effect. In addition, knockdown of DPP8 or DPP9 significantly impairs adipocyte differentiation in preadipocytes. We further uncovered that blocking the expression or activities of DPP8 and DPP9 attenuates PPARγ2 induction during preadipocyte differentiation. Addition of PPARγ agonist thiazolidinediones (TZDs), or ectopic expression of PPARγ2, is able to rescue the adipogenic defect caused by DPP8/9 inhibition in preadipocytes. These results indicate the importance of DPP8 and DPP9 on adipogenesis.

No MeSH data available.


Related in: MedlinePlus

TZDs or ectopic PPARγ2 rescues inhibition of DPP8/9 induced adipogenic defects in 3T3-L1 cells.(A) Representative western blot for the expression of PPARγ in stable 3T3-L1 cells transduced with control plasmid (vehicle) or PPARγ2 plasmid (PPARγ2). The blots were cropped, and the full-length blots are presented in the supplementary information. (B) Oil Red O staining of control cells or PPARγ2 overexpressed 3T3-L1 cells, treated vehicle (NC), DMI (DMI), 500 μM non-selective DPP4 family inhibitor P32/98 (DMI+P32/98) or 20 μM DPP8/9 inhibitor 1G244 (DMI+1G244) at day 8 of differentiation. (C) Oil Red O staining of 3T3-L1 cells treated with 20μM DPP8/9 inhibitor 1G244 (DMI+1G244) or 1G244 plus 1 μM rosiglitazone(DMI+1G244+Rosi) or 5 μM troglitazone (DMI+1G244+Tro). (D) Adipocyte markers, FABP4, adiponectin (AdipoQ) and leptin, were measured in these cells by real time PCR at day 8 of differentiation. β-actin expression was used as an internal control.
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f5: TZDs or ectopic PPARγ2 rescues inhibition of DPP8/9 induced adipogenic defects in 3T3-L1 cells.(A) Representative western blot for the expression of PPARγ in stable 3T3-L1 cells transduced with control plasmid (vehicle) or PPARγ2 plasmid (PPARγ2). The blots were cropped, and the full-length blots are presented in the supplementary information. (B) Oil Red O staining of control cells or PPARγ2 overexpressed 3T3-L1 cells, treated vehicle (NC), DMI (DMI), 500 μM non-selective DPP4 family inhibitor P32/98 (DMI+P32/98) or 20 μM DPP8/9 inhibitor 1G244 (DMI+1G244) at day 8 of differentiation. (C) Oil Red O staining of 3T3-L1 cells treated with 20μM DPP8/9 inhibitor 1G244 (DMI+1G244) or 1G244 plus 1 μM rosiglitazone(DMI+1G244+Rosi) or 5 μM troglitazone (DMI+1G244+Tro). (D) Adipocyte markers, FABP4, adiponectin (AdipoQ) and leptin, were measured in these cells by real time PCR at day 8 of differentiation. β-actin expression was used as an internal control.

Mentions: To determine whether PPARγ2 can rescue the adipogenic defect caused by DPP8/9 inhibition, we generated stable PPARγ2 transduced 3T3-L1 cells (Fig. 5A). Ectopic expression of PPARγ2 was able to rescue P32/98 or 1G244 caused adipogenic defect in preadipocytes (Fig. 5B). The TZDs, rosiglitazone or troglitazone, are known ligands of PPARγ. To investigate whether rosiglitazone or troglitazone were able to rescue the adipogenic defect in DPP8/9 inactivated preadipocytes, we added the DPP8/9 inhibitor 1G244 with either rosiglitazone or troglitazone during 3T3-L1 differentiation. In the absence of rosiglitazone or troglitazone, 1G244 markedly inhibited the adipognesis in 3T3-L1 cells. However, administration of the rosiglitazone or troglitazone completely rescued the adipognesis defect caused by 1G244, as assessed by oil red O staining (Fig. 5C) and confirmed by the expression of adipocyte markers FABP4, adiponectin and leptin (Fig. 5D).


Inhibition of dipeptidyl peptidase 8/9 impairs preadipocyte differentiation.

Han R, Wang X, Bachovchin W, Zukowska Z, Osborn JW - Sci Rep (2015)

TZDs or ectopic PPARγ2 rescues inhibition of DPP8/9 induced adipogenic defects in 3T3-L1 cells.(A) Representative western blot for the expression of PPARγ in stable 3T3-L1 cells transduced with control plasmid (vehicle) or PPARγ2 plasmid (PPARγ2). The blots were cropped, and the full-length blots are presented in the supplementary information. (B) Oil Red O staining of control cells or PPARγ2 overexpressed 3T3-L1 cells, treated vehicle (NC), DMI (DMI), 500 μM non-selective DPP4 family inhibitor P32/98 (DMI+P32/98) or 20 μM DPP8/9 inhibitor 1G244 (DMI+1G244) at day 8 of differentiation. (C) Oil Red O staining of 3T3-L1 cells treated with 20μM DPP8/9 inhibitor 1G244 (DMI+1G244) or 1G244 plus 1 μM rosiglitazone(DMI+1G244+Rosi) or 5 μM troglitazone (DMI+1G244+Tro). (D) Adipocyte markers, FABP4, adiponectin (AdipoQ) and leptin, were measured in these cells by real time PCR at day 8 of differentiation. β-actin expression was used as an internal control.
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Related In: Results  -  Collection

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f5: TZDs or ectopic PPARγ2 rescues inhibition of DPP8/9 induced adipogenic defects in 3T3-L1 cells.(A) Representative western blot for the expression of PPARγ in stable 3T3-L1 cells transduced with control plasmid (vehicle) or PPARγ2 plasmid (PPARγ2). The blots were cropped, and the full-length blots are presented in the supplementary information. (B) Oil Red O staining of control cells or PPARγ2 overexpressed 3T3-L1 cells, treated vehicle (NC), DMI (DMI), 500 μM non-selective DPP4 family inhibitor P32/98 (DMI+P32/98) or 20 μM DPP8/9 inhibitor 1G244 (DMI+1G244) at day 8 of differentiation. (C) Oil Red O staining of 3T3-L1 cells treated with 20μM DPP8/9 inhibitor 1G244 (DMI+1G244) or 1G244 plus 1 μM rosiglitazone(DMI+1G244+Rosi) or 5 μM troglitazone (DMI+1G244+Tro). (D) Adipocyte markers, FABP4, adiponectin (AdipoQ) and leptin, were measured in these cells by real time PCR at day 8 of differentiation. β-actin expression was used as an internal control.
Mentions: To determine whether PPARγ2 can rescue the adipogenic defect caused by DPP8/9 inhibition, we generated stable PPARγ2 transduced 3T3-L1 cells (Fig. 5A). Ectopic expression of PPARγ2 was able to rescue P32/98 or 1G244 caused adipogenic defect in preadipocytes (Fig. 5B). The TZDs, rosiglitazone or troglitazone, are known ligands of PPARγ. To investigate whether rosiglitazone or troglitazone were able to rescue the adipogenic defect in DPP8/9 inactivated preadipocytes, we added the DPP8/9 inhibitor 1G244 with either rosiglitazone or troglitazone during 3T3-L1 differentiation. In the absence of rosiglitazone or troglitazone, 1G244 markedly inhibited the adipognesis in 3T3-L1 cells. However, administration of the rosiglitazone or troglitazone completely rescued the adipognesis defect caused by 1G244, as assessed by oil red O staining (Fig. 5C) and confirmed by the expression of adipocyte markers FABP4, adiponectin and leptin (Fig. 5D).

Bottom Line: Here we demonstrate that the DPP8/9 selective inhibitor 1G244 blocks adipogenesis in preadipocyte 3T3-L1 and 3T3-F422A, while DPP4 and FAP inhibitors have no effect.In addition, knockdown of DPP8 or DPP9 significantly impairs adipocyte differentiation in preadipocytes.These results indicate the importance of DPP8 and DPP9 on adipogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, USA.

ABSTRACT
Adipocytes are the primary cells in adipose tissue, and adipocyte dysfunction causes lipodystrophy, obesity and diabetes. The dipeptidyl peptidase (DPP) 4 family includes four enzymes, DPP4, DPP8, DPP9 and fibroblast activation protein (FAP). DPP4 family inhibitors have been used for the treatment of type 2 diabetes patients, but their role in adipocyte formation are poorly understood. Here we demonstrate that the DPP8/9 selective inhibitor 1G244 blocks adipogenesis in preadipocyte 3T3-L1 and 3T3-F422A, while DPP4 and FAP inhibitors have no effect. In addition, knockdown of DPP8 or DPP9 significantly impairs adipocyte differentiation in preadipocytes. We further uncovered that blocking the expression or activities of DPP8 and DPP9 attenuates PPARγ2 induction during preadipocyte differentiation. Addition of PPARγ agonist thiazolidinediones (TZDs), or ectopic expression of PPARγ2, is able to rescue the adipogenic defect caused by DPP8/9 inhibition in preadipocytes. These results indicate the importance of DPP8 and DPP9 on adipogenesis.

No MeSH data available.


Related in: MedlinePlus