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Inhibition of matrix metalloproteinases and inducible nitric oxide synthase by andrographolide in human osteoarthritic chondrocytes

  • Original Article
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Modern Rheumatology

Abstract

Objective

The aim of this study was to investigate the effects of andrographolide on matrix metalloproteinases (MMP) 1, 3, and 13 and inducible nitric oxide synthase (iNOS) in human articular chondrocytes from osteoarthritic cartilage.

Methods

Passaged chondrocytes were pretreated with or without andrographolide for 2 h, followed by coincubation with interleukin-1 beta (IL-1β) 1 ng/ml for 24 h. Expression levels of MMP-1, 3, and 13, tissue inhibitor of metalloproteinase-1 (TIMP-1), and iNOS were evaluated using real-time-quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting. Nitric oxide (NO) was analyzed using the Griess reaction assay. Involvement of nuclear factor kappa B (NF-κB) was assessed by Western blotting, transient transfection, and luciferase reporter assay.

Results

Andrographolide tested in these in vitro studies was found be an effective antiarthritic agent, as evidenced by potent inhibition of MMP-1, 3, and 13 and iNOS expression, as well as upregulation of TIMP-1 in IL-1β-stimulated human articular chondrocytes (p < 0.05). The mechanism of andrographolide’s inhibitory effects was mediated by attenuating the activation of NF-κB in human chondrocytes in the presence of IL-1β.

Conclusions

Andrographolide was a potent inhibitor of the production of inflammatory and catabolic mediators by chondrocytes, suggesting that this natural compound may merit consideration as a therapeutic agent for treating and preventing osteoarthritis.

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Abbreviations

OA:

Osteoarthritis

IL-1β:

Interleukin-1 beta

MMPs:

Matrix metalloproteinases

TIMP-1:

Tissue inhibitor of metalloproteinases-1

iNOS:

Inducible nitric oxide synthase

NO:

Nitric oxide

NF-κB:

Nuclear factor kappa B

IκB:

Inhibitory kappa B

IKK-β:

Inhibitory kappa B kinase beta

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Acknowledgments

This study was supported by a grant from the Natural Science Foundation of China (30901531), Natural Science Grants of Zhejiang Province (Y207216) and National Natural Science Foundation of China (81071492).

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Department of Orthopedic Surgery, The Second Affiliated Hospital of School of Medicine, Zhejiang University, Jie Fang Road 88#, 310009, Hangzhou, People’s Republic of China

    Qian-hai Ding, Ye Cheng, Yi-ying Qi & **ang-hua Wang

  2. Department of Gastroenterology, The Second Affiliated Hospital of School of Medicine, Zhejiang University, Jie Fang Road 88#, 310009, Hangzhou, People’s Republic of China

    **ao-wei Ji

  3. Department of Surgery, The Second Affiliated Hospital of School of Medicine, Zhejiang University, Jie Fang Road 88#, 310009, Hangzhou, People’s Republic of China

    Yuan-quan Yu

Authors
  1. Qian-hai Ding
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  2. **ao-wei Ji
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  3. Ye Cheng
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  4. Yuan-quan Yu
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  5. Yi-ying Qi
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Corresponding author

Correspondence to **ang-hua Wang.

Electronic supplementary material

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10165_2012_807_MOESM1_ESM.tif

Effects of andrographolide on interleukin (IL)-1β-induced COL2A1, COL9A1, ACAN, ADAMTS-4, 5, and TIMP-3 gene expression. Chondrocytes were pretreated with various andrographolide concentrations for 2 h, followed by stimulation with IL-1β (1 ng/ml) for 24 h. Relative gene expressions were determined by real-time quantitative PCR. Data are expressed as mean ± standard deviatiion (SD). *p < 0.05 andrographolide treated chondrocytes compared with chondrocytes stimulated with IL-1β alone. (TIFF 222 kb)

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Ding, Qh., Ji, Xw., Cheng, Y. et al. Inhibition of matrix metalloproteinases and inducible nitric oxide synthase by andrographolide in human osteoarthritic chondrocytes. Mod Rheumatol 23, 1124–1132 (2013). https://doi.org/10.1007/s10165-012-0807-6

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  • DOI: https://doi.org/10.1007/s10165-012-0807-6

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