Nuclear translocation of STAT3 by in vitro metreleptin administration causes lipolysis in human primary adipocytes

We utilized subcutaneous (SC)- and omental (OM)-derived human primary adipocytes (hPA) from obese male, and investigated whether synthetic analog of leptin, metreleptin, may regulate lipolysis via translocation of STAT3 to the nucleus. We observed that 50 ng/mL of metreleptin increases STAT3 phosphorylation in both SC- and OM-derived hPA. Importantly, we found for the first time that metreleptin is capable of trans-locating STAT3 to the nucleus and STAT3 blockade inhibits metreleptin-induced lipolysis. Our initial data provide novel insights into the role of STAT3 as probable mediator of the action of metreleptin in regulating metabolism.

metreleptin; human primary adipocyte; STAT3; differentiation; lipolysis

Authorship

Seung Kug Choi

Laboratory of Metabolic Engineering, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaLaboratory of Metabolic Engineering, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, South Korea

Institute of Animal Molecular Biotechnology, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaInstitute of Animal Molecular Biotechnology, Korea University, South Korea

Sunmi Park

Institute of Animal Molecular Biotechnology, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaInstitute of Animal Molecular Biotechnology, Korea University, South Korea

Hyun-Seuk Moon

Institute of Animal Molecular Biotechnology, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaInstitute of Animal Molecular Biotechnology, Korea University, South Korea

Biomedical Research Center of Guro Hospital, Research Driven Hospital, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaBiomedical Research Center of Guro Hospital, Research Driven Hospital, Korea University, South Korea

DECLARATION OF INTEREST The authors have declared no conflicts of interest.

AUTHOR CONTRIBUTION All authors participated in the study design, performance and coordination. All authors contributed to writing the manuscript. All authors read and approved the final manuscript.

SCIMAGO INSTITUTIONS RANKINGS

Institute of Animal Molecular Biotechnology, Korea University, South KoreaKorea UniversitySouth KoreaSouth KoreaInstitute of Animal Molecular Biotechnology, Korea University, South Korea

Figures

Figures (2)

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Figure 1-
Regulation of cell differentiation and lipolysis byin vitrometreleptin administration in hPA. Cell culture was performed as described in Methods. (A-B) Cells were treated with metreleptin at indicated concentration for 14 and 28 days, respectively, and cell differentiation was measured by Oil-Red O staining as described in Methods. (C-D) Cells were treated with metreleptin (50 ng/mL) for 12 h and 24 h, respectively, and lipolysis assay was performed as described in Methods. Isoproterenol (10 µM for 2 h) was used as a positive control. All data were analyzed using one-way ANOVA followed by a post hoc test for multiple comparisons. Values are means ± SD; n=3; p<0.05 for any given superscript letter vs all others. C, control.

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Figure 2-
Regulation of STAT3 activation byin vitrometreleptin administration in hPA. Cell culture was performed as described in Methods. (A-C) Cells were treated with metreleptin (50 ng/mL) at indicated time period, and Western Blotting was performed as described in Methods. (D) Cells were treated with metreleptin (50 ng/mL) for 24 h in the absence or presence of STAT3 inhibitor (AG490 for 4 h), and lipolysis assay was performed as described in Methods. All data were analyzed using one-way ANOVA followed by a post hoc test for multiple comparisons. Values are means ± SD; n=3; p<0.05 for any given superscript letter vs all others. C, control.

Figure 1- Regulation of cell differentiation and lipolysis byin vitrometreleptin administration in hPA. Cell culture was performed as described in Methods. (A-B) Cells were treated with metreleptin at indicated concentration for 14 and 28 days, respectively, and cell differentiation was measured by Oil-Red O staining as described in Methods. (C-D) Cells were treated with metreleptin (50 ng/mL) for 12 h and 24 h, respectively, and lipolysis assay was performed as described in Methods. Isoproterenol (10 µM for 2 h) was used as a positive control. All data were analyzed using one-way ANOVA followed by a post hoc test for multiple comparisons. Values are means ± SD; n=3; p<0.05 for any given superscript letter vs all others. C, control.

Figure 2- Regulation of STAT3 activation byin vitrometreleptin administration in hPA. Cell culture was performed as described in Methods. (A-C) Cells were treated with metreleptin (50 ng/mL) at indicated time period, and Western Blotting was performed as described in Methods. (D) Cells were treated with metreleptin (50 ng/mL) for 24 h in the absence or presence of STAT3 inhibitor (AG490 for 4 h), and lipolysis assay was performed as described in Methods. All data were analyzed using one-way ANOVA followed by a post hoc test for multiple comparisons. Values are means ± SD; n=3; p<0.05 for any given superscript letter vs all others. C, control.

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Nuclear translocation of STAT3 by in vitro metreleptin administration causes lipolysis in human primary adipocytes

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[1] DECLARATION OF INTEREST The authors have declared no conflicts of interest.