TRPV4 regulates migration and tube formation of human retinal capillary endothelial cells

BMC Ophthalmol. 2018 Feb 12;18(1):38. doi: 10.1186/s12886-018-0697-2.

Abstract

Background: Ca2+ entry plays an important role in modulating endothelial cell migration and tube formation. Transient receptor potential cation channel subfamily V member 4 (TRPV4) is a Ca2+-permeable channel that is widely expressed in endothelial cells. It has been reported that TRPV4 is expressed in HRCECs and regulates Ca2+ entry. However, the function of TRPV4 in human retinal capillary endothelial cells (HRCECs) remains unknown.

Methods: In this study we used western blot and immunostaining assay to verify TRPV4 expression in HRCECs. And then we pretreated HRCECs with HC067047 and transfected with specific shRNA of TRPV4. The functional presence of TrpV4 was determined by using fluorescence, migration and tube formation assay in TrpV4 knockdown cells or control cells.

Results: Using western blot and immunostaining, we confirmed TRPV4 expression in HRCECs. Moreover, inhibition of TRPV4 using the specific inhibitor HC067047 and the knockdown of TRPV4 with shRNA significantly suppressed tube formation and migration by HRCECs.

Conclusions: TRPV4 is essential for HRCEC migration and tube formation, and maybe a potential therapeutic target for retinal vascular diseases.

Keywords: HRCEC; Migration; TRPV4; Tube formation.

MeSH terms

  • Blotting, Western
  • Calcium / metabolism
  • Calcium Channel Agonists / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Cell Count
  • Cell Movement / physiology*
  • Endothelial Cells / cytology*
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism*
  • Fluorescent Antibody Technique, Indirect
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics
  • RNA, Small Interfering / genetics
  • Retinal Vessels / physiology*
  • TRPV Cation Channels / physiology*
  • Transfection

Substances

  • Calcium Channel Agonists
  • Calcium Channel Blockers
  • RNA, Small Interfering
  • TRPV Cation Channels
  • TRPV4 protein, human
  • Calcium