The di-peptide Trp-His activates AMP-activated protein kinase and enhances glucose uptake independently of insulin in L6 myotubes

摘要

class="kwd-title">Abbreviations: AICAR, 5-aminoimidazole-4-carboxamide ribonucleoside; AMPK, AMP-activated protein kinase; CaMK, Ca²⁺/calmodulin-dependent kinase; DMEM, Dulbecco’s modified Eagle’s medium; 2DG, 2-deoxy-d-glucose; Glut, glucose transporter; IRS-1, insulin receptor substrate-1; KRH, Krebs–Ringer–HEPES buffer; PHT1, peptide/histidine transporter 1; PM, plasma membrene; TEA, triethanolamine; VDCC, voltage-dependent calcium channel class="kwd-title">Keywords: AMP-activated protein kinase, Trp-His, LKB1, Glucose transporter 4, Peptide transporter class="head no_bottom_margin" id="idm139725925773840title">AbstractThe di-peptide Trp-His (WH) has vasorelaxant and anti-atherosclerotic functions. We hypothesized that WH has multiple biological functions and may aid AMP-activated protein kinase (AMPK) activation and affect the glucose transport system in skeletal muscle.First, we examined whether WH or His-Trp (HW) can activate AMPKα. Treatment of L6 myotubes with WH or HW significantly increased phosphorylation of AMPKα. WH activated AMPK independently of insulin and significantly increased glucose uptake into L6 myotubes following translocation of glucose transporter 4 (Glut4) to the plasma membrane. This activation was induced by the LKB1 pathway but was independent of changes in intracellular Ca²⁺ levels and the Ca²⁺/calmodulin-dependent kinase pathway. L6 myotubes express only one type of oligopeptide transporter, peptide/histidine transporter 1 (PHT1, also known as SLC15a4), and WH is incorporated into cells and activates AMPKα following PHT1-mediated cell uptake.These findings indicate that (1) WH activates AMPK and insulin independently enhances glucose uptake following translocation of Glut4 to the plasma membrane, (2) activation of AMPKα by WH is mediated by the LKB1 pathway, without altering the Ca²⁺-dependent pathway, and (3) L6 myotubes express only one type of peptide transporter (PHT1; SLC15a4), which incorporates WH into cells to activate AMPKα.