Impaired wound healing in hypoxic renal tubular cells: roles of hypoxia-inducible factor-1 and glycogen synthase kinase 3β/β-catenin signaling.
Wound and subsequent healing are frequently associated with hypoxia. Although hypoxia induces angiogenesis for tissue remodeling during wound healing, it may also affect the healing response of parenchymal cells. Whether and how wound healing is affected by hypoxia in kidney cells and tissues is currently unknown. Here, we used scratch-wound healing and transwell migration models to examine the effect of hypoxia in cultured renal proximal tubular cells (RPTC). Wound healing and migration were significantly slower in hypoxic (1% oxygen) RPTC than normoxic (21% oxygen) cells. Hypoxia-inducible factor-1α (HIF-1α) was induced during scratch-wound healing in normoxia, and the induction was more evident in hypoxia. Nevertheless, HIF-1α-null and wild-type cells healed similarly after scratch wounding. Moreover, activation of HIF-1α with dimethyloxalylglycine in normoxic cells did not suppress wound healing, negating a major role of HIF-1α in wound healing in this model. Scratch-wound healing was also associated with glycogen synthase kinase 3β (GSK3β)/β-catenin signaling, which was further enhanced by hypoxia. Pharmacological inhibition of GSK3β resulted in β-catenin expression, accompanied by the suppression of wound healing and transwell cell migration. Ectopic expression of β-catenin in normoxic cells could also suppress wound healing, mimicking the effect of hypoxia. Conversely, inhibition of β-catenin via dominant negative mutants or short hairpin RNA improved wound healing and transwell migration in hypoxic cells. The results suggest that GSK3β/β-catenin signaling may contribute to defective wound healing in hypoxic renal cells and tissues.
Department of Cellular Biology and Anatomy, Georgia Health Sciences University, Augusta, GA 30912, USA.
Jianping
J
Peng
Ganesan
G
Ramesh
Lin
L
Sun
Zheng
Z
Dong
D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology*; D002465:Cell Movement; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000502:physiology*; D000071679:Glycogen Synthase Kinase 3 beta; D051793:Hypoxia-Inducible Factor 1 / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D007687:Kidney Tubules, Proximal / Q000502:physiology*; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology*; D051176:beta Catenin / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D051793:Hypoxia-Inducible Factor 1; D051176:beta Catenin; D012313:RNA; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; C000605508:Gsk3b protein, rat; D038362:Glycogen Synthase Kinase 3