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Protein Tyrosine Phosphatase-1B (PTP-1B) Knockdown Improves Palmitate-Induced Insulin Resistance in C2C12 Skeletal Muscle Cells

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Lipids

Abstract

Insulin resistance is the central defect in type 2 diabetes and obesity. During the development of insulin resistance a lipid accumulation is accompanied by increased PTP-1B expression in the muscle. The aim of this study was to examine the effects of PTP-1B knockdown on insulin signaling and insulin resistance in the presence or absence of palmitate in C2C12 skeletal muscle cells. A stable C2C12 cell line was established using short hairpin RNA (shRNA) to knockdown protein expression of PTP1B. Analysis of PTP-1B protein expression and phosphorylation and protein levels of IRS-1 and Akt were detected by western blot. The effects of PTP-1B knockdown on the glucose uptake was also measured in C2C12 cells. The stable C2C12 cell line harboring the PTP-1B shRNA showed 62% decrease in the PTP-1B protein levels. 0.5 mM palmitate significantly induced insulin resistance in both control (26%) and PTP-1B knockdown cells (16.5%) compared to the untreated cells. Under treatment with palmitate, insulin stimulated phosphorylation of IRS-1 (Tyr632) and Akt (Ser473) in knockdown cells was significantly 1.55- and 1.86-fold, respectively, greater than the controls. In the presence of palmitate, insulin dependent glucose uptake was significantly about 3-fold higher in PTP-1B knockdown stable C2C12 cells compared to the control cells. Our data showed that decreasing the PTP-1B protein level by shRNA can enhance the activity of important elements of insulin signaling. The improvement in insulin action persisted even in palmitate treated insulin resistant myotubes.

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Abbreviations

BSA:

Bovine serum albumin

2-DOG:

2-Deoxyglucose

IR:

Insulin receptor

IRS-1:

Insulin receptor substrate-1

LAR:

Leukocyte antigen related phosphatase

PTP-1B:

Protein tyrosine phosphatase-1B

ShRNA:

Small hairpin RNA

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Acknowledgments

This work was supported by a Grant from the Iran National Science Foundation (INSF) (grant no. 86062/23).

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. Iran

    Salar Bakhtiyari & Mohammad Taghikhani

  2. Department of Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, I.R. Iran

    Reza Meshkani

  3. Endocrinology and Metabolism Research Centre, Shariati Hospital, Tehran University of Medical Sciences, Tehran, I.R. Iran

    Bagher Larijani

  4. Division of Clinical Biochemistry, Department of Pediatric Laboratory Medicine, Hospital for Sick Children, University of Toronto, Ontario, Canada

    Khosrow Adeli

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  1. Salar Bakhtiyari
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  2. Reza Meshkani
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Correspondence to Reza Meshkani.

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Bakhtiyari, S., Meshkani, R., Taghikhani, M. et al. Protein Tyrosine Phosphatase-1B (PTP-1B) Knockdown Improves Palmitate-Induced Insulin Resistance in C2C12 Skeletal Muscle Cells. Lipids 45, 237–244 (2010). https://doi.org/10.1007/s11745-010-3394-3

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  • DOI: https://doi.org/10.1007/s11745-010-3394-3

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