Expression analysis of three isoforms of hyaluronan synthase and hyaluronidase in the synovium of knees in osteoarthritis and rheumatoid arthritis by quantitative real-time reverse transcriptase polymerase chain reaction

Abstract

Hyaluronan is a major molecule in joint fluid and plays a crucial role in joint motion and the maintenance of joint homeostasis. The concentration and average molecular weight of hyaluronan in the joint fluids are reduced in osteoarthritis and rheumatoid arthritis. To elucidate the underlying mechanism, we analyzed the message expression of three isoforms of hyaluronan synthase and hyaluronidase from knee synovium, using real-time reverse transcriptase polymerase chain reaction. Synovia were obtained from 17 patients with osteoarthritis, 14 patients with rheumatoid arthritis, and 20 healthy control donors. The message expression of hyaluronan synthase-1 and -2 in the synovium of both types of arthritis was significantly less than in the control synovium, whereas that of hyaluronidase-2 in the synovium of both arthritides was significantly greater than in the control synovium. The decreased expression of the messages for hyaluronan synthase-1 and -2 and/or the increased expression of the message for hyaluronidase-2 may be reflected in the reduced concentration and decreased average molecular weight of hyaluronan in the joint fluids of patients with osteoarthritis and rheumatoid arthritis.

Figure 1

Relative expression levels of the messages for hyaluronan synthase-1, -2, and -3 and hyaluronidase-1, -2, and -3 in the synovium of knees in osteoarthritis (OA) and rheumatoid arthritis (RA). Total RNA was isolated from knee synovium and expression levels of the messages were relatively quantified by real-time reverse transcriptase polymerase chain reaction. The expression levels of the messages in OA and RA are expressed by longitudinal bars relative to those of the control value expressed as 1.0. The lines outside the bars represent the standard deviation of the change in threshold cycle numbers (ΔC_T) corrected with C_T values of β-actin message used as an internal standard. * P < 0.01.