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Role for sterol regulatory element binding protein-1c activation in mediating skeletal muscle insulin resistance via repression of rat insulin receptor substrate-1 transcription

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Abstract

Aims/hypothesis

Sterol regulatory element binding protein-1c (SREBP-1c) is a master regulator of fatty acid synthase and controls lipogenesis. IRS-1 is the key insulin signalling mediator in skeletal muscle. In the present study, we investigated the role of SREBP-1c in the regulation of IRS-1 in skeletal muscle cells.

Methods

L6 muscle cells were treated with palmitic acid (PA) or metformin. Adenovirus vectors expressing Srebp-1c (also known as Srebf1) and small interfering RNA (siRNA) against Srebp-1c were transfected into the L6 cells. Protein–DNA interactions were assessed by luciferase reporter analysis, electrophoretic mobility shift assay and chromatin immunoprecipitation assay.

Results

We found that both gene and protein expression of SREBP-1c was increased in contrast to IRS-1 expression in PA-treated L6 cells. SREBP-1c overproduction decreased Irs-1 mRNA and IRS-1 protein expression in a dose-dependent manner, and suppressed the resultant insulin signalling, whereas SERBP-1c knockdown by Serbp-1c siRNA blocked the downregulation of IRS-1 induced by PA. Protein–DNA interaction studies demonstrated that SREBP-1c was able to bind to the rat Irs-1 promoter region, thereby repressing its gene transcription. Of particular importance, we found that metformin treatment downregulated Srebp-1c promoter activity, decreased the specific binding of SREBP-1c to Irs-1 promoter and upregulated Irs-1 promoter activity in PA-cultured L6 cells.

Conclusions/interpretation

Our data indicate for the first time that SREBP-1c activation participates in skeletal muscle insulin resistance through a direct effect of suppressing Irs-1 transcription. These findings imply that SREBP-1c could serve as an attractive therapeutic target for insulin resistance.

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Abbreviations

2-NBDG:

2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-amino]-2-deoxy-d-glucose

AMPK:

AMP-activated protein kinase

bHLH:

Basic helix-loop-helix

ChIP:

Chromatin immunoprecipitation assay

EMSA:

Electrophoretic mobility shift assay

FAS:

Fatty acid synthase

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GFP:

Green fluorescent protein

LXR:

Liver X receptor

PA:

Palmitic acid

PI3K:

Phosphatidlyinositol-3-kinase

SD rat:

Sprague-Dawley rat

siRNA:

Small interfering RNA

SRE:

Sterol regulatory element

SREBP-1c:

Sterol regulatory element binding protein-1c

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Acknowledgements

We would like to thank Prof. Li Xiaoying from Shanghai Ruijin Hospital for his constructive discussion during manuscript preparation. Some of the data were presented as an abstract at the 49th Annual European Association for the Study of Diabetes (EASD) Meeting in 2013 (number 2194).

Funding

This work was sponsored by grants from the National Natural Science Foundation of China Grant Award (81270906, 30800539), 973 project (2012CB517506), National Science Fund for Distinguished Young Scholars (81025005), China postdoctoral Science Foundation (2012M521050), Jiangsu postdoctoral Science Foundation, Jiangsu Province’s Key Provincial Talents Program (RC2011011), Jiangsu Province’s Key Discipline of Medicine (XK201105), the Key Project of Nanjing Medical Science and Technology Development Foundation (ZKX11017), National Natural Science Foundation of China Grant Award (81000338, 81070636), New Drug Development, Construction and management of Clinical Biobank for Major Disease (2011ZX0907-001-08), the Project of National Key Clinical Division, Jiangsu Natural Science Foundation (KA037) and Guangdong Natural Science Foundation (10151008901000033).

Contribution statement

YB contributed to the study design, data interpretation, drafting the article, and final approval of the version to be published. WW contributed to the acquisition of data, drafting the article and approval of the final version. JS contributed to the study design, acquisition of data, drafting the article and approval of the final version. HL contributed the study design, data analysis, drafting the article and approval of the final version. WY, YC, ST, SC, MC, and SS contributed to acquisition of data, drafting the article and approval of the final version. QG contributed to the acquisition of data, drafting the article and approval of the final version. DZ contributed to the study design, acquisition of data, revision of the manuscript and final approval of the version to be published. JW contributed to the study design, acquisition of data, revision of the manuscript and final approval of the version to be published.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

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Authors and Affiliations

  1. Department of Endocrinology, Drum Tower hospital affiliated to Nanjing University Medical School, No321 Zhongshan Road, Nanjing, 210008, People’s Republic of China

    Yan Bi, Yingying Chen, Sunyinyan Tang, Shanmei Shen & Dalong Zhu

  2. Department of Endocrinology, Drum Tower Clinical Medical College of Nanjing Medical University, Nanjing, 210008, People’s Republic of China

    Yan Bi, Wenwen Yin, Shu Cao & Dalong Zhu

  3. Department of Endocrinology, Wuxi People’s hospital of Nanjing Medical University, Wuxi, People’s Republic of China

    Wenjun Wu

  4. Department of Oncology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, People’s Republic of China

    Junfeng Shi

  5. Department of Endocrinology, the 3rd affiliated hospital of Sun Yat-sen University, No 600 Tianhe Road, Guangzhou, 510630, People’s Republic of China

    Hua Liang, Mengyin Cai & Jianping Weng

  6. Key Lab of Diabetology of Guangdong Province, Guangzhou, People’s Republic of China

    Jianping Weng

  7. Nanjing University Medical School, Nanjing, People’s Republic of China

    Qian Gao

Authors
  1. Yan Bi
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  2. Wenjun Wu
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  3. Junfeng Shi
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  4. Hua Liang
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  6. Yingying Chen
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  7. Sunyinyan Tang
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  8. Shu Cao
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  9. Mengyin Cai
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  10. Shanmei Shen
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  11. Qian Gao
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  12. Jianping Weng
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  13. Dalong Zhu
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Corresponding authors

Correspondence to Yan Bi, Jianping Weng or Dalong Zhu.

Additional information

Yan Bi, Wenjun Wu, Junfeng Shi and Hua Liang contributed equally to this study.

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Bi, Y., Wu, W., Shi, J. et al. Role for sterol regulatory element binding protein-1c activation in mediating skeletal muscle insulin resistance via repression of rat insulin receptor substrate-1 transcription. Diabetologia 57, 592–602 (2014). https://doi.org/10.1007/s00125-013-3136-1

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