Role of tumour necrosis factor-a in the regulation of T-type calcium channel current in HL-1 cells

Clin Exp Pharmacol Physiol. 2016 Jul;43(7):706-11. doi: 10.1111/1440-1681.12585.

Abstract

Increasing evidence indicates that inflammation contributes to the initiation and perpetuation of atrial fibrillation (AF). Although tumour necrosis factor (TNF)-α levels are increased in patients with AF, the role of TNF-α in the pathogenesis of AF remains unclear. Besides L-type Ca(2+) currents (IC a,L ), T-type Ca(2+) currents (IC a,T ) also plays an important role in the pathogenesis of AF. This study was designed to use the whole-cell voltage-clamp technique and biochemical assays to explore if TNF-α is involved in the pathogenesis of AF through regulating IC a,T in atrial myocytes. It was found that compared with sinus rhythm (SR) controls, T-type calcium channel (TCC) subunit mRNA levels were decreased, while TNF-α expression levels were increased, in human atrial tissue from patients with AF. In murine atrial myocyte HL-1 cells, after culturing for 24 h, 12.5, 25 and 50 ng/mL TNF-α significantly reduced the protein expression levels of the TCC α1G subunit in a concentration-dependent manner. The peak current was reduced by the application of 12.5 or 25 ng/mL TNF-α in a concentration-dependent manner (from -15.08 ± 1.11 pA/pF in controls to -11.89 ± 0.83 pA/pF and -8.54 ± 1.55 pA/pF in 12.5 or 25 ng/mL TNF-α group respectively). TNF-α application also inhibited voltage-dependent inactivation of IC a,T, shifted the inactivation curve to the left. These results suggest that TNF-α is involved in the pathogenesis of AF, probably via decreasing IC a,T current density in atrium-derived myocytes through impaired channel function and down-regulation of channel protein expression. This pathway thus represents a potential pathogenic mechanism in AF.

Keywords: HL-1 cells; T-type Ca2+ channel; atrial fibrillation; tumour necrosis factor-α.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Atrial Fibrillation / metabolism
  • Calcium Channels, L-Type / metabolism
  • Calcium Channels, T-Type / metabolism*
  • Cell Line
  • Down-Regulation / physiology
  • Female
  • Heart Atria / metabolism
  • Humans
  • Male
  • Mice
  • Middle Aged
  • Myocytes, Cardiac / metabolism
  • Patch-Clamp Techniques / methods
  • Tumor Necrosis Factor-alpha / metabolism*

Substances

  • Calcium Channels, L-Type
  • Calcium Channels, T-Type
  • Tumor Necrosis Factor-alpha