Effect of oleanolic acid on the activity, secretion and gene expression of matrix metalloproteinase-3 in articular chondrocytes in vitro and the production of matrix metalloproteinase-3 in vivo

  • Kang, Dong-Geun (Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University) ;
  • Lee, Hyun Jae (Department of Health Management, Sahmyook University) ;
  • Kim, Kun Tae (Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University) ;
  • Hwang, Sun-Chul (Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University) ;
  • Lee, Choong Jae (Department of Pharmacology, School of Medicine, Chungnam National University) ;
  • Park, Jin Sung (Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University)
  • Received : 2016.11.03
  • Accepted : 2016.12.01
  • Published : 2017.03.01


In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.


Supported by : National Research Foundation of Korea (NRF)


  1. Aigner T, McKenna L. Molecular pathology and pathobiology of osteoarthritic cartilage. Cell Mol Life Sci. 2002;59:5-18.
  2. Mankin HJ. The response of articular cartilage to mechanical injury. J Bone Joint Surg Am. 1982;64:460-466.
  3. Dean DD, Martel-Pelletier J, Pelletier JP, Howell DS, Woessner JF Jr. Evidence for metalloproteinase and metalloproteinase inhibitor imbalance in human osteoarthritic cartilage. J Clin Invest. 1989;84:678-685.
  4. Kullich W, Fagerer N, Schwann H. Effect of the NSAID nimesulide on the radical scavenger glutathione S-transferase in patients with osteoarthritis of the knee. Curr Med Res Opin. 2007;23:1981-1986.
  5. Birkedal-Hansen H, Moore WG, Bodden MK, Windsor LJ, Birkedal-Hansen B, DeCarlo A, Engler JA. Matrix metalloproteinases: a review. Crit Rev Oral Biol Med. 1993;4:197-250.
  6. Burrage PS, Mix KS, Brinckerhoff CE. Matrix metalloproteinases: role in arthritis. Front Biosci. 2006;11:529-543.
  7. Garnero P, Rousseau JC, Delmas PD. Molecular basis and clinical use of biochemical markers of bone, cartilage, and synovium in joint diseases. Arthritis Rheum. 2000;43:953-968.<953::AID-ANR1>3.0.CO;2-Q
  8. Lin PM, Chen CT, Torzilli PA. Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage. Osteoarthritis Cartilage. 2004;12:485-496.
  9. Freemont AJ, Hampson V, Tilman R, Goupille P, Taiwo Y, Hoyland JA. Gene expression of matrix metalloproteinases 1, 3, and 9 by chondrocytes in osteoarthritic human knee articular cartilage is zone and grade specific. Ann Rheum Dis. 1997;56:542-549.
  10. Goupille P, Jayson MI, Valat JP, Freemont AJ. Matrix metalloproteinases: the clue to intervertebral disc degeneration? Spine (Phila Pa 1976). 1998;23:1612-1626.
  11. Kanyama M, Kuboki T, Kojima S, Fujisawa T, Hattori T, Takigawa M, Yamashita A. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids of patients with temporomandibular joint osteoarthritis. J Orofac Pain. 2000;14:20-30.
  12. Jo H, Park JS, Kim EM, Jung MY, Lee SH, Seong SC, Park SC, Kim HJ, Lee MC. The in vitro effects of dehydroepiandrosterone on human osteoarthritic chondrocytes. Osteoarthritis Cartilage. 2003;11:585-594.
  13. Little CB, Barai A, Burkhardt D, Smith SM, Fosang AJ, Werb Z, Shah M, Thompson EW. Matrix metalloproteinase 13-deficient mice are resistant to osteoarthritic cartilage erosion but not chondrocyte hypertrophy or osteophyte development. Arthritis Rheum. 2009;60:3723-3733.
  14. Neuhold LA, Killar L, Zhao W, Sung ML, Warner L, Kulik J, Turner J, Wu W, Billinghurst C, Meijers T, Poole AR, Babij P, DeGennaro LJ. Postnatal expression in hyaline cartilage of constitutively active human collagenase-3 (MMP-13) induces osteoarthritis in mice. J Clin Invest. 2001;107:35-44.
  15. Yoshihara Y, Nakamura H, Obata K, Yamada H, Hayakawa T, Fujikawa K, Okada Y. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids from patients with rheumatoid arthritis or osteoarthritis. Ann Rheum Dis. 2000;59:455-461.
  16. Echtermeyer F, Bertrand J, Dreier R, Meinecke I, Neugebauer K, Fuerst M, Lee YJ, Song YW, Herzog C, Theilmeier G, Pap T. Syndecan-4 regulates ADAMTS-5 activation and cartilage breakdown in osteoarthritis. Nat Med. 2009;15:1072-1076.
  17. Stanton H, Rogerson FM, East CJ, Golub SB, Lawlor KE, Meeker CT, Little CB, Last K, Farmer PJ, Campbell IK, Fourie AM, Fosang AJ. ADAMTS5 is the major aggrecanase in mouse cartilage in vivo and in vitro. Nature. 2005;434:648-652.
  18. Nam DC, Kim BK, Lee HJ, Shin HD, Lee CJ, Hwang SC. Effects of prunetin on the proteolytic activity, secretion and gene expression of MMP-3 in vitro and production of MMP-3 in vivo. Korean J Physiol Pharmacol. 2016;20:221-228.
  19. Park JS, Kim DK, Shin HD, Lee HJ, Jo HS, Jeong JH, Choi YL, Lee CJ, Hwang SC. Apigenin regulates interleukin-$1{\beta}$-induced production of matrix metalloproteinase both in the knee joint of rat and in primary cultured articular chondrocytes. Biomol Ther (Seoul). 2016;24:163-170.
  20. Park JS, Lee HJ, Lee DY, Jo HS, Jeong JH, Kim DH, Nam DC, Lee CJ, Hwang SC. Chondroprotective effects of wogonin in experimental models of osteoarthritis in vitro and in vivo. Biomol Ther (Seoul). 2015;23:442-448.
  21. Kang BJ, Ryu J, Lee CJ, Hwang SC. Luteolin inhibits the activity, secretion and gene expression of MMP-3 in cultured articular chondrocytes and production of MMP-3 in the rat knee. Biomol Ther (Seoul). 2014;22:239-245.
  22. Kashyap D, Sharma A, Tuli HS, Punia S, Sharma AK. Ursolic acid and oleanolic acid: pentacyclic terpenoids with promising antiinflammatory activities. Recent Pat Inflamm Allergy Drug Discov. 2016;10:21-33.
  23. Tsao SM, Yin MC. Antioxidative and antiinflammatory activities of asiatic acid, glycyrrhizic acid, and oleanolic acid in human bronchial epithelial cells. J Agric Food Chem. 2015;63:3196-3204.
  24. Jeon SJ, Lee HJ, Lee HE, Park SJ, Gwon Y, Kim H, Zhang J, Shin CY, Kim DH, Ryu JH. Oleanolic acid ameliorates cognitive dysfunction caused by cholinergic blockade via TrkB-dependent BDNF signaling. Neuropharmacology. 2016;113:100-109.
  25. Long C, Yang J, Yang H, Li X, Wang G. Attenuation of renal ischemia/reperfusion injury by oleanolic acid preconditioning via its antioxidant, anti-inflammatory, and anti-apoptotic activities. Mol Med Rep. 2016;13:4697-4704.
  26. Choi JK, Kim SW, Kim DS, Lee JY, Lee S, Oh HM, Ha YS, Yoo J, Park PH, Shin TY, Kwon TK, Rho MC, Kim SH. Oleanolic acid acetate inhibits rheumatoid arthritis by modulating T cell immune responses and matrix-degrading enzymes. Toxicol Appl Pharmacol. 2016;290:1-9.
  27. Moon PD, Jeong HS, Chun CS, Kim HM. Baekjeolyusin-tang and its active component berberine block the release of collagen and proteoglycan from IL-$1{\beta}$-stimulated rabbit cartilage and downregulate matrix metalloproteinases in rabbit chondrocytes. Phytother Res. 2011;25:844-850.
  28. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst. 1990;82:1107-1112.
  29. Bonnet CS, Walsh DA. Osteoarthritis, angiogenesis and inflammation. Rheumatology (Oxford). 2005;44:7-16.
  30. Goldring MB, Otero M, Tsuchimochi K, Ijiri K, Li Y. Defining the roles of inflammatory and anabolic cytokines in cartilage metabolism. Ann Rheum Dis. 2008;67 Suppl 3:iii75-82.
  31. Kobayashi M, Squires GR, Mousa A, Tanzer M, Zukor DJ, Antoniou J, Feige U, Poole AR. Role of interleukin-1 and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage. Arthritis Rheum. 2005;52:128-135.
  32. Loeser RF. Molecular mechanisms of cartilage destruction: mechanics, inflammatory mediators, and aging collide. Arthritis Rheum. 2006;54:1357-1360.
  33. Aida Y, Maeno M, Suzuki N, Shiratsuchi H, Motohashi M, Matsumura H. The effect of IL-1beta on the expression of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in human chondrocytes. Life Sci. 2005;77:3210-3221.
  34. Pantsulaia I, Kalichman L, Kobyliansky E. Association between radiographic hand osteoarthritis and RANKL, OPG and inflammatory markers. Osteoarthritis Cartilage. 2010;18:1448-1453.
  35. Lijnen HR. Matrix metalloproteinases and cellular fibrinolytic activity. Biochemistry (Mosc). 2002;67:92-98.
  36. Zhou K, Hu L, Liao W, Yin D, Rui F. Coptisine prevented IL-${\beta}$-induced expression of inflammatory mediators in chondrocytes. Inflammation. 2016;39:1558-1565.
  37. Lu S, Xiao X, Cheng M. Matrine inhibits IL-$1{\beta}$-induced expression of matrix metalloproteinases by suppressing the activation of MAPK and $NF-{\kappa}B$ in human chondrocytes in vitro. Int J Clin Exp Pathol. 2015;8:4764-4772.
  38. Piao T, Ma Z, Li X, Liu J. Taraxasterol inhibits IL-$1{\beta}$-induced inflammatory response in human osteoarthritic chondrocytes. Eur J Pharmacol. 2015;756:38-42.
  39. Jeong JW, Lee HH, Choi EO, Lee KW, Kim KY, Kim SG, Hong SH, Kim GY, Park C, Kim HK, Choi YW, Choi YH. Schisandrae fructus inhibits IL-$1{\beta}$-induced matrix metalloproteinases and inflammatory mediators production in SW1353 human chondrocytes by suppressing $NF-{\kappa}B$ and MAPK activation. Drug Dev Res. 2015;76:474-483.
  40. Kang GD, Lim S, Kim DH. Oleanolic acid ameliorates dextran sodium sulfate-induced colitis in mice by restoring the balance of Th17/Treg cells and inhibiting $NF-{\kappa}B$ signaling pathway. Int Immunopharmacol. 2015;29:393-400.