Effect of GCSB-5, a Herbal Formulation, on Monosodium Iodoacetate-Induced Osteoarthritis in Rats

Joon-Ki Kim, Sang-Won Park, Jung-Woo Kang, Yu-Jin Kim, Sung Youl Lee, Joonshik Shin, Sangho Lee, Sun-Mee Lee, Joon-Ki Kim, Sang-Won Park, Jung-Woo Kang, Yu-Jin Kim, Sung Youl Lee, Joonshik Shin, Sangho Lee, Sun-Mee Lee

Abstract

Therapeutic effects of GCSB-5 on osteoarthritis were measured by the amount of glycosaminoglycan in rabbit articular cartilage explants in vitro, in experimental osteoarthritis induced by intra-articular injection of monoiodoacetate in rats in vivo. GCSB-5 was orally administered for 28 days. In vitro, GCSB-5 inhibited proteoglycan degradation. GCSB-5 significantly suppressed the histological changes in monoiodoacetate-induced osteoarthritis. Matrix metalloproteinase (MMP) activity, as well as, the levels of serum tumor necrosis factor-α, cyclooxygenase-2, inducible nitric oxide synthase protein, and mRNA expressions were attenuated by GCSB-5, whereas the level of interleukin-10 was potentiated. By GCSB-5, the level of nuclear factor-κB p65 protein expression was significantly attenuated but, on the other hand, the level of inhibitor of κB-α protein expression was increased. These results indicate that GCSB-5 is a potential therapeutic agent for the protection of articular cartilage against progression of osteoarthritis through inhibition of MMPs activity, inflammatory mediators, and NF-κB activation.

Figures

Figure 1
Figure 1
GAG release in rabbit articular cartilage explant cultures at 24, 48, and 72 h. Rabbit articular cartilage explants were stimulated with rhIL-1α (5 ng/mL). The amount of GAG release stimulated by rhIL-1α (∘) increased approximately 3.6 times compared to control (●) at 72 h. GCSB-5 (1.0 × 10−3 (▼) and 1.0 × 10−2 (∆) mg/mL) and diclofenac (30 μM (□)) efficiently inhibited the GAG release. However, a high concentration of GCSB-5 (1.0 × 10−1 (■) mg/mL) slightly inhibited it. Each value represents the mean ± S.E.M. from 6 articular cartilage explants cultures per group. **Significantly different (P < 0.01) from control. + and ++Significantly different (P < 0.05, P < 0.01) from rhIL-1α.
Figure 2
Figure 2
Roentgenography and histopathological features of osteoarthritic lesion in the knee joint of rats 28 days after intra-articular injection of MIA (H and E staining, ×100; SOFG staining, ×100). Control (a) represents intact normal joint feature. Vehicle-treated MIA (b) shows a severely damaged joint with rough edges around the tibia and femur, indicative of bone lysis, swelling, and tendency of patellar displacement. However, these damages were reduced significantly by treatment with 600 mg/kg GCSB-5 (c) and 5 mg/kg diclofenac (d). SOGF-stained control (e) represents normal cartilage PG staining, whereas vehicle-treated MIA (f) represents severely damaged cartilage showing marked fibrillation and the depletion of SOFG staining with separation of cartilage from subchondral bone. 600 mg/kg GCSB-5 (g) and 5 mg/kg diclofenac (h) treatments significantly reduced cartilage damage. H and E stained control (i) represents the normal status of joint cartilage, whereas vehicle-treated MIA (j) represents severely damaged cartilage showing widespread cell necrosis and inflammation. However, treatment with 600 mg/kg GCSB-5 (k) and 5 mg/kg diclofenac (l) treatments significantly reduced joint cartilage damage.
Figure 3
Figure 3
Activities of MMP-2 (Gelatinase A) and MMP-9 (Gelatinase B) assessed by zymography in knee joint cartilages obtained 28 days after MIA injection. The latent and active amounts of gelatinase were combined to give a total value for each gelatinase. Each value represents the mean ± S.E.M. from 6 rats per group. aSignificantly different (P < 0.01) from control. bSignificantly different (P < 0.05) from vehicle-treated MIA.
Figure 4
Figure 4
TNF-α, IL-1β, and IL-10 mRNA expressions in cartilage from knee joints of rats at day 2 after MIA injection. Each value represents the mean ± S.E.M. from 6 rats per group. a,bSignificantly different (P < 0.01, P < 0.05) from control. c,dSignificantly different (P < 0.01, P < 0.05) from vehicle-treated MIA.
Figure 5
Figure 5
COX-2 and iNOS protein expressions in cartilage from knee joints of rats at day 2 after MIA injection. Each value represents the mean ± S.E.M. from 6 rats per group. a,bSignificantly different (P < 0.01, P < 0.05) from control. c,dSignificantly different (P < 0.01, P < 0.05) from vehicle-treated MIA.
Figure 6
Figure 6
COX-2 and iNOS mRNA expressions in cartilage from knee joints of rats at day 2 after MIA injection. Each value represents the mean ± S.E.M. from 6 rats per group. aSignificantly different (P < 0.05) from control. bSignificantly different (P < 0.05) from vehicle-treated MIA.
Figure 7
Figure 7
Nuclear NF-κB p65 and cytosolic IκB-α protein expressions in cartilage from knee joints of rats at day 2 after MIA injection. Each value represents the mean ± S.E.M. from 6 rats per group. a,bSignificantly different (P < 0.05, P < 0.01) from control. c,dSignificantly different (P < 0.05, P < 0.01) from vehicle-treated MIA.

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Source: PubMed

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