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:: Volume 15, Issue 4 (12-2013) ::
2013, 15(4): 387-394 Back to browse issues page
Investigating the Effects of TNF-α knockdown Gene on Insulin Resistance in C2C12 Muscle Cells in the Presence and Absence of Palmitate
Somayeh Pashaei , Salar Bakhtiyari Dr , Karimeh Haghani Dr, Faraj allah Maleki
, bakhtiyari-s@medilam.ac.ir
Abstract:   (4941 Views)

Introduction: Insulin resistance plays a major role in type 2 diabetes and obesity. In this disorder, lipid accumulation is accompanied by increased TNF-α expression in the muscle. The aims of this study were to evaluate the effects of tumor necrosis factor-α (TNF-α) gene knockdown on key elements of insulin signaling pathway (IRS-1 and Akt) and insulin resistance in C2C12 muscle cells in the presence and absence of palmitate. Materials and Methods: To knockdown protein expression of TNF-α, the C2C12 cells were transfected with the shRNA containing antisense sequence of murine TNF-α gene. The analysis of TNF-α protein expression and phosphorylation and protein levels of IRS-1and Akt were subsequently detected by western blot. Results: In TNF-α knockdown cells, the protein expression level of TNF-α was reduced by 58%. Under treatment with palmitate, insulin stimulated phosphorylation of IRS-1 (Tyr632) and Akt (Ser473) in knockdown cells was increased 1.7 and 2.6 fold, respectively, compared to the controls. Conclusions: Our findings showed that decreasing the TNF-α protein level can enhance the activity of the important elements of insulin signaling pathway (IRS-1 and Akt), leading to the improvement of insulin resistance in myotubes, data suggesting that TNF-α may potentially be a therapeutic target for fatty acid induced insulin resistance observed in type 2 diabetes and metabolic syndrome.

 

Keywords: Tumor necrosis factor- alpha, Short hairpin RNA interference, Insulin resistance, Knockdown, Palmitate
Full-Text [PDF 336 kb]   (2221 Downloads)    
Type of Study: Original | Subject: Biochemistry
Received: 2013/02/25 | Accepted: 2013/05/15 | Published: 2013/12/15
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Pashaei S, Bakhtiyari S, Haghani K, Maleki F A. Investigating the Effects of TNF-α knockdown Gene on Insulin Resistance in C2C12 Muscle Cells in the Presence and Absence of Palmitate. Iranian Journal of Endocrinology and Metabolism. 2013; 15 (4) :387-394
URL: http://ijem.sbmu.ac.ir/article-1-1522-en.html


Volume 15, Issue 4 (12-2013) Back to browse issues page
مجله ی غدد درون‌ریز و متابولیسم ایران، دو ماهنامه  پژوهشی مرکز تحقیقات غدد درون‌ریز و متابولیسم، Iranian Journal of Endocrinology and Metabolism
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