Induction of osteogenic protein-1 expression by interleukin-1beta in cultured rabbit articular chondrocytes.
To elucidate the effects of interleukin-1beta (IL-1beta) on osteogenic protein-1 (OP-1) gene expression in a polylayer culture of rabbit articular chondrocytes, we measured rabbit OP-1 mRNA using quantitative TaqMan reverse transcriptase-polymerase chain reaction (RT-PCR) techniques. Rabbit articular chondrocytes were isolated and cultured in minimum essential medium eagle alpha modification containing 10% fetal bovine serum for 7 days. IL-1beta was then added and cultures were continued for 48 or 96 hours. OP-1 gene expression was detected in cell cultures both with and without addition of IL-1beta. However, the level of expression was very low in the control group. OP-1 gene expression was significantly increased about 450- to 800-fold in IL-1beta-treated groups (0.1, 1, and 10 ng/ml) versus the control group. Evaluation of serial changes in OP-1 expression after addition of IL-1beta (10 ng/ml) revealed that OP-1 gene expression increased rapidly after addition of IL-1beta, reaching a peak at 48 hours, and then decreasing. Simultaneous assay of CD44 expression demonstrated a rapid increase, similar to that of OP-1 expression, following addition of IL-1beta: this was followed by a more gradual increase. Assay of hyaluronan synthase-2 (HAS-2) expression following addition of IL-1beta showed an increase after OP-1 expression had already reached a peak. Our results demonstrate that OP-1 expression is induced by IL-1beta and suggest that this expression, like that of HAS-2, may play a role as a protective mechanism against inflammatory cytokines.
Yoshida
Satoshi
S
Department of Biochemistry, Iwate Medical University, Morioka, Japan. satto@fj8.so-net.ne.jp
Kubota
Yoshiko
Y
Toba
Tamotsu
T
Horiuchi
Saburo
S
Shimamura
Tadashi
T
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D002358:Cartilage, Articular / Q000166:cytology / Q000502:physiology*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000502:physiology*; D015870:Gene Expression / Q000187:drug effects*; D014453:Glucuronosyltransferase / Q000235:genetics; D018960:Hyaluronan Receptors / Q000235:genetics; D007375:Interleukin-1 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D016212:Transforming Growth Factor beta
D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D018960:Hyaluronan Receptors; D007375:Interleukin-1; D016212:Transforming Growth Factor beta; D014453:Glucuronosyltransferase