J Rheum Dis.  2018 Jul;25(3):188-196. 10.4078/jrd.2018.25.3.188.

Compound K Inhibits Interleukin-1β-induced Expression of Inflammatory Mediators and Matrix Metalloproteinases by Inhibiting Mitogen-activated Protein Kinase Activation in Chondrocytes

• Park EH
• Kim JS
• Lee JS
• Lee YJ
• Song YW
• Lee EY

• KMID: 2415607
• DOI: http://doi.org/10.4078/jrd.2018.25.3.188

Abstract

OBJECTIVE
This study examined the anti-inflammatory and chondroprotective effects of compound K (CK), a ginsenoside metabolite, on chondrocytes from osteoarthritis (OA) patients following stimulation with interleukin (IL)-1β.
METHODS
Articular cartilage samples were obtained from six OA patients undergoing total knee replacement surgery. Nitric oxide (NO) production was measured by the Griess reaction. Subsequently, the mRNA and protein levels of matrix metalloproteinases (MMPs), inducible NO synthase (iNOS), and mitogen-activated protein kinases (MAPKs) were examined by a reverse transcription-polymerase chain reaction and western blot analysis. Cartilage degradation was assessed using a glycosaminoglycan (GAG) assay.
RESULTS
CK inhibited IL-1β-induced NO production and iNOS expression in a dose-dependent manner. In addition, it inhibited the IL-1 β-stimulated release of MMP-1, -3, and -13 and tissue inhibitor of matrix metalloproteinase-1 from OA patient chondrocytes. In addition, CK effectively suppressed the IL-1β-induced phosphorylation of p38, extracellular signal-regulated kinase-1/2, and c-Jun N-terminal kinase MAPKs. Moreover, the IL-1β-mediated release of GAG was inhibited by CK in a dose-dependent manner.
CONCLUSION
CK inhibited the IL-1β-induced expression of inflammatory mediators and MMPs by, at least in part, inhibiting MAPK activation, and has potential as a therapeutic agent for the treatment of OA.

Keyword

Panax; Ginsenosides; Nitric oxide; Matrix metalloproteinases; Osteoarthritis

MeSH Terms

Arthroplasty, Replacement, Knee
Blotting, Western
Cartilage
Cartilage, Articular
Chondrocytes*
Ginsenosides
Humans
Interleukin-1
Interleukins
JNK Mitogen-Activated Protein Kinases
Matrix Metalloproteinase 1
Matrix Metalloproteinases*
Mitogen-Activated Protein Kinases
Nitric Oxide
Nitric Oxide Synthase
Osteoarthritis
Panax
Phosphorylation
Protein Kinases*
RNA, Messenger
Ginsenosides
Interleukin-1
Interleukins
JNK Mitogen-Activated Protein Kinases
Matrix Metalloproteinase 1
Matrix Metalloproteinases
Mitogen-Activated Protein Kinases
Nitric Oxide
Nitric Oxide Synthase
Protein Kinases
RNA, Messenger

Figure

• 

  Supplementary Figure 1. Chemical structure of (A) compound K (CK) and (B) ginsenoside Rb1.

• 

  Figure 1. Interleukin (IL)-1β-induced nitric oxide (NO) production inhibited by (A) compound K (CK) and (B) Rb1. (C) Strong inhibitory effect of CK on NO production is related to decreased expression of induced NO synthase (iNOS), (D) but the expression of iNOS was not inhibited by Rb1. The values presented are the means±standard error of mean. of three independent experiments.*p＜0.05, **p＜0.005.

• 

  Figure 2. Effects of compound K (CK) and Rb1 on interleukin (IL)-1β-induced mRNA expression of induced nitric oxide synthase (iNOS), matrix metalloproteinases (MMPs), and tissue inhibitor of metalloproteinases-1 (TIMP-1). The cells were pretreated with different concentrations of CK or Rb1 for 1 hour before incubation with IL-1β (2 ng/mL). Following 24 hours of treatment, total RNA was isolated, and reverse tran-scription-polymerase chain reaction was performed using iNOS, MMP-1, MMP-3, MMP-13 and TIMP-1 primers.

• 

  Figure 3. Compound K (CK) inhibits interleukin (IL)-1β-induced mitogen-activated protein kinase activation. ERK: extracellular signal-regulated kinase, JNK: c-Jun N-terminal kinase.

• 

  Figure 4. Activity of matrix metalloproteinases (MMPs) was inhibited by (A) compound K (CK) on the zymographic study. Production of MMP-1, MMP-3, MMP-13, and tissue inhibitor of metalloproteinases-1 (TIMP-1) measured by enzyme-linked immunosorbent assay was inhibited by (B) CK in a dose-dependent manner but not by (C) Rb1. IL: interleukin. The values presented are the means±standard error of mean of three independent experiments. *p＜0.05, **p＜0.005.

• 

  Figure 5. Effect of (A) compound K (CK) and (B) Rb1 on chondrocyte degeneration analyzed through the glycosaminoglycan (GAG) assay. IL: interleukin. The values presented are the means±standard error of mean of three independent experiments. *p＜0.05, **p＜0.005.

• 

  Figure 6. Effects of (A) compound K (CK) and (B) Rb1 on the cell viability of chondrocytes. The cells were cultured with different concentrations of CK (0∼10 μM) or Rb1 (0∼200 μM) for 24 hours. The cell viability was determined by MTT assay. IL: interleukin. The values are means±standard error of mean of three independent experiments.

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