title: Hypercholesterolemia inhibits re-endothelialization of arterial injuries by TRPC channel activation.
abstract: OBJECTIVE
  After arterial injury, endothelial cell (EC) migration is essential for healing, but lipid oxidation products activate TRPC6 and TRPC5 ion channels, leading to increased intracellular calcium and inhibition of EC migration in vitro. The objective of this study was to further evaluate the role of TRPC channels in EC migration in vitro and to validate in vitro findings in an in vivo model.
  
  
  METHODS
  Mouse aortic ECs were cultured, and the effect of lysophosphatidylcholine, the major lysophospholipid in oxidized low-density lipoprotein, on migration was assessed in a razor-scrape assay. EC healing after a carotid injury with electrocautery was evaluated in wild-type (WT), TRPC6(-/-), and TRPC5(-/-) mice receiving either a chow or high-cholesterol (HC) diet.
  
  
  RESULTS
  Lysophosphatidylcholine inhibited EC migration of WT ECs to 22% of baseline and of TRPC5(-/-) ECs to 53% of baseline but had minimal effect on TRPC6(-/-) EC migration. Hypercholesterolemia severely impaired EC healing in vivo, with 51.4% ± 1.8% and 24.9% ± 2.0% of the injury resurfaced with ECs at 5 days in chow-fed and HC-fed WT mice, respectively (P < .001). Hypercholesterolemia did not impair healing in TRPC6(-/-) mice, with coverage of 48.4% ± 3.4% and 46.8% ± 1.6% in chow-fed and HC-fed TRPC6(-/-) mice, respectively. Hypercholesterolemia had a reduced inhibitory effect in TRPC5(-/-) mice, with EC coverage of 51.7% ± 3.0% and 37.% ± 1.4% in chow-fed and HC-fed TRPC5(-/-) mice, respectively.
  
  
  CONCLUSIONS
  Results suggest that activation of TRPC6 and TRPC5 channels is the key contributor to impaired endothelial healing of arterial injuries in hypercholesterolemic mice.
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, Cleveland, Ohio; Department of Vascular Surgery, Cleveland Clinic, Cleveland, Ohio.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Rosenbaum
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, Cleveland, Ohio.
authors.forename: Pinaki
authors.initials: P
authors.lastname: Chaudhuri
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, Cleveland, Ohio; Department of Vascular Surgery, Cleveland Clinic, Cleveland, Ohio. Electronic address: grahamL@ccf.org.
authors.forename: Linda M
authors.initials: LM
authors.lastname: Graham
mesh_terms: D000818:Animals; D001158:Arteries / Q000293:injuries*; D015415:Biomarkers / Q000097:blood; D002118:Calcium / Q000032:analysis; D002465:Cell Movement / Q000502:physiology; D042783:Endothelial Cells / Q000502:physiology; D004730:Endothelium, Vascular / Q000502:physiology*; D006937:Hypercholesterolemia / Q000097:blood / Q000503:physiopathology* / Q000652:urine; D066298:In Vitro Techniques; D007249:Inflammation / Q000097:blood; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D051379:Mice; D018384:Oxidative Stress; D050052:TRPC Cation Channels / Q000502:physiology*; D014945:Wound Healing / Q000502:physiology
chemical_list: D015415:Biomarkers; D008244:Lysophosphatidylcholines; D050052:TRPC Cation Channels; D002118:Calcium