High-fat feeding increases insulin receptor and IRS-1 coimmunoprecipitation with SOCS-3, IKKalpha/beta phosphorylation and decreases PI-3 kinase activity in muscle

Abstract

Suppressor of cytokine signaling (SOCS) proteins and/or activation of the proinflammatory pathway have been postulated as possible mechanisms that may contribute to skeletal muscle insulin resistance. Thus, the aims of the present investigation were to determine in high-fat-fed skeletal muscle: 1) whether SOCS-3 protein concentration is increased, 2) whether coimmunoprecipitation of SOCS-3 with the insulin receptor-beta subunit and/or IRS-1 is increased, and 3) whether select components of the proinflammatory pathway are altered. Thirty-two male Sprague-Dawley rats were assigned to either control (CON, n = 16) or high-fat-fed (HF, n = 16) dietary groups for 12 wk and then subjected to hind limb perfusions in the presence (n = 8/group) or absence (n = 8/group) of insulin. Insulin-stimulated skeletal muscle 3-MG transport rates and PI-3 kinase activity were greater (P < 0.05) in CON. IRS-1 tyrosine phosphorylation was decreased (P < 0.05), and IRS-1 serine 307 phosphorylation was increased (P < 0.05) in HF. Insulin receptor-beta (IR-beta) subunit coimmunoprecipitation with IRS-1 was reduced in HF. SOCS-3 protein concentration and SOCS-3 coimmunoprecipitation with both the IR-beta subunit and IRS-1 was increased (P < 0.05) in HF. IKKalpha/beta serine phosphorylation was increased (P < 0.05), IkappaBalpha protein concentration was decreased (P < 0.05) and IkappaBalpha serine phosphorylation was increased (P < 0.05) in HF. Increased colocalization of SOCS-3 with both the IR-beta subunit and IRS-1 may provide steric hindrance that prevents IRS-1 from interacting with IR-beta, while increased IKKbeta serine phosphorylation may contribute to increasing IRS-1 serine phosphorylation, both of which independently can have deleterious effects on insulin-stimulated PI-3 kinase activation in high-fat-fed rodent skeletal muscle.

Fig. 1.

TNF-α protein concentration (A), IKKα/β protein concentration (B), IKKα/β serine phosphorylation (pS) (C), and IκBα protein concentration and IκBα serine phosphorylation (pS) (D) in skeletal muscle obtained from control (CON) and high-fat fed (HF) animals. *Significantly different from CON (P < 0.05). Values are expressed as means ± SE.

Fig. 2.

SOCS-3 protein concentration (A), and SOCS-3 coimmunoprecipitation (B) with insulin receptor β-subunit and SOCS-3 coimmunoprecipitation with IRS-1 in skeletal muscle obtained from CON and HF animals. *Significantly different from CON (P < 0.05). Values are expressed as means ± SE.