J Rheum Dis.  2015 Jun;22(3):154-166. 10.4078/jrd.2015.22.3.154.

Compound K, a Metabolite of Ginsenosides, Attenuates Collagen-induced Arthritis in Mice

• Lee YJ
• Song KY
• Lee EY
• Kang HS
• Song YW

• KMID: 2222881
• DOI: http://doi.org/10.4078/jrd.2015.22.3.154

Abstract

OBJECTIVE
Although several ginsenosides have been reported to have anti-arthritic activity, few in vivo studies of the anti-arthritic effects of compound K (CK), a major metabolite of ginsenosides, have been conducted. Therefore, we investigated the preventative and therapeutic effects of CK on collagen-induced arthritis (CIA).
METHODS
CK was administered to CIA mice preventively and therapeutically and post-treatment bone microarchitectural characteristics, histopathological changes, and serum levels of anti-collagen antibodies, tumor necrosis factor-alpha, and interleukin (IL)-17 were investigated. We also examined cytokine production by type II collagen (CII)-stimulated splenocytes and mRNA expression of matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinase (TIMP)-1, receptor activator of nuclear factor-kappaB ligand (RANKL), and osteoprotegerin (OPG) in the joint tissues.
RESULTS
CK reduced the severity of CIA preventively and therapeutically (all p<0.05). Additionally, CK dose-dependently decreased histopathological signs of arthritis and improved microarchitectural characteristics (all p<0.05) at 10 to 20 mg/kg/d in CIA mice. CK treatment significantly decreased the serum levels of anti-CII immunoglobulin G (p<0.01) and the secretion of interferon-gamma and IL-2 from stimulated splenocytes (all p<0.05). Furthermore, MMP-3/TIMP-1 and RANKL/OPG ratios were suppressed in CK treated mice (all p<0.01).
CONCLUSION
CK attenuated CIA via suppression of the humoral immune response and modulation of joint-destructive mediators. These results suggest that CK has therapeutic potential in rheumatoid arthritis.

Keyword

Panax; Ginsenoside M1; Experimental arthritis; Rheumatoid arthritis

MeSH Terms

Animals
Antibodies, Neoplasm
Arthritis
Arthritis, Experimental*
Arthritis, Rheumatoid
Collagen Type II
Ginsenosides*
Immunity, Humoral
Immunoglobulin G
Interferon-gamma
Interleukin-2
Interleukins
Joints
Matrix Metalloproteinases
Mice*
Necrosis
Osteoprotegerin
Panax
RANK Ligand
RNA, Messenger
Antibodies, Neoplasm
Collagen Type II
Ginsenosides
Immunoglobulin G
Interferon-gamma
Interleukin-2
Interleukins
Matrix Metalloproteinases
Osteoprotegerin
RANK Ligand
RNA, Messenger

Figure

• 

  Figure 1. The effect of compound K (CK) on collagen-induced arthritis (CIA) disease activity. CK (0, 5, 10, or 20 mg/kg/d) was administered 1 day (preventive trial) or 14 days (therapeutic trial) after the boosting immunization of type II collagen (A). Preventive (B, n=9) and therapeutic (C, n=9) administration of CK significantly decreased arthritis scores along the disease course. However, the suppressive effects of CK were not dose-dependent in the dose range tested (5 to 20 mg/kg/d). Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test.

• 

  Figure 2. Representative sections of joint tissues (hindfoot and knee joints) stained with hematoxylin-eosin (H&E) and Masson's Trichrome (MT) in therapeutic models (scale bar, 50 μ m; A). Control group showed typical findings of collagen-induced arthritis (CIA); severe inflammatory cells infiltration, synovial hypertrophy, joint space narrowing, and bone and cartilage damage. These arthritic findings were decreased in mice therapeutically administered with compound K (CK). The histologic arthritis scores in knee joints significantly decreased with increasing dose of CK (B, n=6). Error bars represent the standard error of the mean. † p-val-ue was calculated by Jonckheere's trend test.

• 

  Figure 3. Microarchitectural analysis using micro-computed tomography (micro-CT) in therapeutic models (n=4 in each group, respectively). Representative micro-CT images of the hind paws (A) showed that the erosion in the metatarsophalangeal joints was decreased in compound K (CK)-treated groups. (B, C) Microarchitectural parameters included bone volume/total tissue volume (BV/TV), bone surface/bone volume (BS/BV), cross-sectional thickness (Cs.th), and trabecular thickness (Tb.th). Therapeutic CK administration significantly increased BV/TV, Cs.th, and Tb.th values and significantly decreased BS/BV values. Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test; †p-values were calculated by Kruskal-Wallis test.

• 

  Figure 4. Serum levels of anti-type II collagen (CII) antibodies in therapeutic models. Therapeutical administration of compound K (CK) produced a significant reduction of anti-CII antibody IgG2 a (A) and IgG2 b (B, n=6 in each group). Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test; † p-values were calculated by Kruskal-Wallis test.

• 

  Figure 5. Cytokine production from the type II collagen stimulated splenocytes in therapeutic models. Splenocytes were isolated at the time of sacrifice after 4 weeks of compound K (CK) treatment and were stimulated with bovine CII for 48 h (n=6 in each group). When tumor necrosis factor (TNF)-α (A), interleukin (IL)-2 (B), interferon (IFN)-γ (C), and IL-4 (D) levels were measured in the culture media, IL-2 and IFN-γ were dose-dependently decreased in CK-treated groups. However, no significant effects on TNF-α or IL-4 production were observed. Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test; †p-values were calculated by Kruskal-Wallis test.

• 

  Figure 6. Serum levels of tumor necrosis factor (TNF)-α and interleukin (IL)-17 in therapeutic models (n=6 in each group). Therapeutic compound K (CK) administration did not significantly affect TNF-α (A) and IL-17 (B) levels. Error bars represent the standard error of the mean.

• 

  Figure 7. Effects of compound K (CK) on matrix metalloproteinase (MMP)-3 and MMP-13 mRNA expression in the hind feet of therapeutic models. When MMP-3 and MMP-13 mRNA levels in the hind foot tissue were analyzed using quantitative real-time polymerase chain reaction (n=6 in each group), therapeutic administration of CK significantly decreased expression of MMP-3 (A) and MMP-13 (B). The suppressive effect of CK on MMP-3 expression remained significant after adjustment for tissue inhibitors of metalloproteinase (TIMP)-1 mRNA (C, n=4 in each group). MMP-13/TIMP-1 ratios (D, n=4 in each group) tended to decrease in a dose dependent manner. Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test; † p-values were calculated by Kruskal-Wallis test.

• 

  Figure 8. Effects of compound K (CK) on receptor activator of nuclear factor-κ B ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in the hind feet of therapeutic models. When RANKL and OPG mRNA levels in the hind foot tissue were analyzed using quantitative real-time polymerase chain reaction (n=6 in each group), therapeutic CK administration significantly decreased RANKL mRNA expression (A) and tended to increase OPG mRNA (B). Consequently, the ratios of RANKL/OPG were significantly reduced in CK administered mice (C). Error bars represent the standard error of the mean. *p＜0.05 by Kruskal-Wallace test with Dunn's multiple comparison test; † p-values were calculated by Kruskal-Wallis test.

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