Journal List > J Korean Rheum Assoc > v.14(3) > 1003561

Lee, Lee, Park, Kim, Cho, Han, Yoo, and Yang: The Suppressive Effect of Rifampicin in Animal Model of Hemophilic Synovitis

Abstract

Objective:

Hemophilic arthropathy, which results from recurrent intra-articular bleeding, is a proliferative synovitis, but the sequence of pathogenic events in hemophilic synovitis (HS) is not known in detail. To investigate the pathogenic mechanism of HS and to evaluate the suppressive effect of rifampicin for HS, we designed this study.

Methods:

Twenty normal male New Zealand white rabbits weighing approximately 2,000 gm were used for this study. We injected 1 mL of autologous whole blood of the rabbits into the right knee joint and normal saline into the left knee joint (control) thrice a week for 10 weeks and sacrificed 10 of them. We injected 10 mg of rifampicin into the right knee joint of HS, which is 5 rabbits of remained 10, once a week from 11th week until 15th week. At 11th week and 20th week, the rabbits were sacrificed and both knee joints of each rabbit were opened, synovial membrane specimens were collected and examined pathologically and biologically such as mRNA of IL-1, TNF-α, MMP-1 and MMP-3 in hemophilic synovium using by real-time quantitative PCR method (comparative Ct method).

Results:

At 20th week, in rifampicin treated group showed decreased proliferative, and infiltrated mononuclear cells compared with control group. And mRNA of TNF-α, IL-1 and MMP-3 levels were decreased also.

Conclusion:

In animal model of HS, histological changes showed the same as human hemophilic synovitis. And this study suggested that rifampicin has a controlling effect on the inflammatory process of HS by suppression of inflammatory cytokine production in the experimental hemophilic synovitis model.

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Fig. 1.
The blood injected knee of rabbit (arrow) was more swollen compared with the other knee.
jkra-14-201f1.tif
Fig. 2.
The blood injected right knee shows degenerative change by X-ray.
jkra-14-201f2.tif
Fig. 3.
The synovium of blood injected knee shows proliferation and fat tissue was increased under synovium (A) normal saline injected knee synovium (B) blood injected knee synovium (H&E stain, x 400).
jkra-14-201f3.tif
Fig. 4.
The expression of TNF-α, IL-1, MMP-1 and MMP-3 mRNA in synovium of animal model of Hemathrosis (HA) by real time-PCR. Control is saline-injected knee, HA is blood-injected synovium without rifampicin, and RFP is blood-injected synovium treated with rifampicin.
jkra-14-201f4.tif
Table 1.
Primer and PCR protocol according to each cytokine
Cytokine Primer PCR cycle Fragment size
IL-1beta 5’-AACCAACAAGTGGTGTTCTC-3’ Denaturation for 40 sec at 94oC
5’-GACTCAAATTCCAGCTTGTC-3 ‘ Annealing for 40 sec at 62oC 223bp
Primer extension for 1 min at 72oC∗35 cycle
TNF-α 5’-CACCACTTAGAAACCTGGAC-3 ‘ Denaturation for 40 sec at 63oC 216bp
5’-TAGGTCTGGGTGACAACTTC-3’
MMP-1 5’-ACCAAAACGAGAGTGCTGCT-3’ Annealing for 40 sec at 61oC 272bp
5’-AAGCTGAAGTTCGGGTGAGA-3’
MMP-3 5 ‘-GCTTTGCTCAGCCTATCCAC-3’ Primer extension for 1 min at 72oC∗35 cycle 282bp
MMP-3 5’-GTGCCAGGGAAGGTACTGAA-3’
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