title: Growth factor regulation of intracellular pH homeostasis under hypoxic conditions in isolated equine articular chondrocytes.
abstract: Hypoxia and acidosis are recognized features of inflammatory arthroses. This study describes the effects of IGF-1 and TGF-β(1) on pH regulatory mechanisms in articular cartilage under hypoxic conditions. Acid efflux, reactive oxygen species (ROS), and mitochondrial membrane potential were measured in equine articular chondrocytes isolated in the presence of serum (10% fetal calf serum), IGF-1 (1, 10, 50, 100 ng/ml) or TGF-β(1) (0.1, 1, 10 ng/ml) and then exposed to a short-term (3 h) hypoxic insult (1% O(2)). Serum and 100 ng/ml IGF-1 but not TGF-β(1) attenuated hypoxic regulation of pH homeostasis. IGF-1 appeared to act through mitochondrial membrane potential stabilization and maintenance of intracellular ROS levels in very low levels of oxygen. Using protein phosphorylation inhibitors PD98059 (25 µM) and wortmannin (200 nM) and Western blotting, ERK1/2 and PI-3 kinase pathways are important for the effect of IGF-1 downstream to ROS generation in normoxia but only PI-3 kinase is implicated in hypoxia. These results show that oxygen and growth factors interact to regulate pH recovery in articular chondrocytes by modulating intracellular oxygen metabolites.
authors.affiliation: Faculty of Health and Life Sciences, Department of Musculoskeletal Biology, University of Liverpool, Leahurst Campus, Neston CH64 7TE, United Kingdom. p.i.milner@liverpool.ac.uk
authors.forename: Peter I
authors.initials: PI
authors.lastname: Milner
authors.forename: Hannah C
authors.initials: HC
authors.lastname: Smith
authors.forename: Rebecca
authors.initials: R
authors.lastname: Robinson
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Wilkins
authors.forename: John S
authors.initials: JS
authors.lastname: Gibson
mesh_terms: D000136:Acid-Base Equilibrium / Q000187:drug effects*; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects*; D019902:Chondrocytes / Q000187:drug effects*; D005419:Flavonoids / Q000494:pharmacology; D006706:Homeostasis / Q000187:drug effects; D006736:Horses; D006863:Hydrogen-Ion Concentration; D000860:Hypoxia / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010100:Oxygen / Q000494:pharmacology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D005419:Flavonoids; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017382:Reactive Oxygen Species; D016212:Transforming Growth Factor beta; D007334:Insulin-Like Growth Factor I; D010100:Oxygen; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D000077191:Wortmannin