Stimulation of hyaluronan metabolism by interleukin-1alpha in human articular cartilage.
OBJECTIVE
To determine the effects of interleukin-1alpha (IL-1alpha) on the expression of hyaluronan synthase (HAS), CD44, and aggrecan in human articular chondrocytes, and to assess the net result of these metabolic changes on the accumulation of hyaluronan within articular cartilage.


METHODS
Normal human articular cartilage slices, as well as isolated chondrocytes, were treated with IL-1alpha. Changes in the relative expression of messenger RNA (mRNA) for HAS-2, CD44, and aggrecan were determined by competitive, quantitative reverse transcriptase-polymerase chain reaction. Hyaluronan accumulation was characterized by staining with a hyaluronan-specific binding protein and by fluorophore-assisted carbohydrate electrophoresis, while proteoglycan content was determined by alcian blue and Safranin O staining, CD44 protein expression by immunohistochemistry, and aggrecan biosynthesis by 35S-sulfate incorporation. Changes in cell-associated matrix sizes were visualized by a particle exclusion assay.


RESULTS
IL-1alpha stimulated the expression of HAS-2 and CD44 mRNA (3.5-fold and 3-fold, respectively), but inhibited the expression of aggrecan mRNA. In IL-1-treated chondrocytes, extracellular hyaluronan decreased, while intracellular accumulation of hyaluronan was enhanced. Together with the decrease in expression of aggrecan, a dramatic reduction in cell-associated matrix was observed. IL-1-treated cartilage slices displayed a prominent depletion of aggrecan as well as hyaluronan within the upper layers of the tissue. The regional loss of hyaluronan coincided with a regional up-regulation of CD44.


CONCLUSIONS
These data demonstrate that IL-1alpha stimulates HAS-2 at the same time as it inhibits the expression of aggrecan. Although hyaluronan biosynthesis is up-regulated, so too is the expression of CD44 and the internalization/catabolism of hyaluronan. The net result is a loss of hyaluronan in areas of the articular cartilage where increases in CD44 expression are most prominent. This depletion of hyaluronan in the upper layers of the tissue likely facilitates the prominent loss of aggrecan from the tissue.
Nishida
Y
Y
Rush Medical College, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612, USA.
D'Souza
A L
AL
Thonar
E J
EJ
Knudson
W
W
D053674:Aggrecans; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D046508:Culture Techniques; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology; D016326:Extracellular Matrix Proteins; D014453:Glucuronosyltransferase / Q000235:genetics; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid / Q000378:metabolism*; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D037181:Lectins, C-Type; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D014018:Tissue Distribution
D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D018960:Hyaluronan Receptors; D007375:Interleukin-1; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D006820:Hyaluronic Acid; D014453:Glucuronosyltransferase; C101754:HAS2 protein, human; D000076002:Hyaluronan Synthases