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Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2018; 82(2); 82-88; doi: 10.1155/2015/329418

In vitro study of matrix metalloproteinases 1, 2, 9, 13 and serum amyloid A mRNAs expression in equine fibroblast-like synoviocytes treated with doxycycline.

Abstract: Application of synthetic matrix metalloproteinases (MMPs) inhibitors, such as doxycycline is one of the possible therapeutic options for osteoarthritis. However, little is known about the protective mechanism of doxycycline in equine models on MMPs inhibitors as well as on serum amyloid A (SAA) gene expression. This study investigated the effects of doxycycline on mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA of equine fibroblast-like synoviocytes (FLSs). The FLSs were established from synovial fluids of clinically normal metacarpophalangeal joints of 6 skeletally mature horses. The cells were treated with either 10 or 100 μg/mL of doxycycline for 48 h. The mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA were assessed using real-time polymerase chain reaction (PCR). Treatment with doxycycline resulted in significantly decreased mRNA expression of MMP-1 in FLSs at both concentrations ( = 0.001). No significant differences were detected among groups for MMP-2, MMP-9, and MMP-13 ( > 0.05). Only a tendency towards a decrease in mRNA expression level of SAA in the presence of doxycycline could be detected. Doxycycline inhibits MMP-1 gene expression at the transcript level. These findings indicate that doxycycline can protect the articular environment through inhibition of MMP-1 at transcript level. L’application d’inhibiteurs synthétiques des métalloprotéinases de la matrice (MMP), telle que la doxycycline, est une des options thérapeutiques possibles pour l’ostéoarthrite. Toutefois, peu de choses sont connues sur le mécanisme protecteur de la doxycycline dans les modèles équins des inhibiteurs des MMP, de même que sur l’expression génique de l’amyloïde sérique A (SAA). La présente étude visait à déterminer les effets de la doxycycline sur l’expression de l’ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA des synoviocytes équins apparentés aux fibroblastes (FLS). Les FLS ont été établis à partir du liquide synovial provenant d’articulations métacarpo-phalangiennes cliniquement normales de six chevaux squelettiquement matures. Les cellules ont été traitées avec 10 ou 100 μg/mL de doxycycline pendant 48 h. L’expression d’ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA a été évaluée par réaction d’amplification en chaîne par la polymérase en temps réel. Le traitement avec la doxycycline a causé une diminution significative de l’expression de MMP-1 par les FLS et ce pour les deux concentrations ( = 0,001). Aucune différence significative ne fut détectée parmi les groupes MMP-2, MMP-9, et MMP-13 ( > 0,05). Seulement une tendance à la diminution de l’expression d’ARNm de SAA en présence de doxycycline pouvait être notée. La doxycycline inhibe l’expression génique de MMP-1 à l’étape de la transcription. Ces informations indiquent que la doxycycline peut protéger l’environnement articulaire en inhibant MMP-1 à l’étape de la transcription.(Traduit par Docteur Serge Messier).
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Publication Date: 2018-05-15 PubMed ID: 29755186PubMed Central: PMC5914083DOI: 10.1155/2015/329418Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates how doxycycline, a matrix metalloproteinases (MMPs) inhibitor, impacts the mRNA expression of MMPs and serum amyloid A in horse cells (specifically equine fibroblast-like synoviocytes). The conclusion suggests that doxycycline reduces mRNA expression of MMP-1 which could potentially assist in alleviating conditions like osteoarthritis in horses.

Methods Utilized

  • Equine fibroblast-like synoviocytes (FLSs) were extracted from the synovial fluids of the metacarpophalangeal joints (part of the horse’s lower skeletal limb) of 6 mature horses that showed no clinical abnormalities.
  • The FLSs were then treated with two different concentrations of doxycycline (10 and 100 μg/mL) for a period of 48 hours.
  • The impacts of the doxycycline treatment on the mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13 and SAA were then assessed through real-time polymerase chain reaction (PCR), which is a specific method used to amplify and quantify nucleic acids (like DNA and RNA).

Key Findings

  • The researchers found a significant decrease in MMP-1 mRNA expressions in FLSs when treated with doxycycline, irrespective of the dose (either 10 or 100 μg/mL).
  • However, when it came to mRNA expression of MMP-2, MMP-9, MMP-13, the doxycycline treatment didn’t show any significant impact.
  • The study also found a mere tendency towards a reduction in SAA mRNA expression when treated with doxycycline, but the change was not significant enough to reach a firm conclusion.

Implications and Conclusions

  • The significant decrease in MMP-1 mRNA expression due to doxycycline suggests that this drug may possess a protective mechanism against osteoarthritis in equine models. This is believed to be achieved by inhibiting MMP-1 at the mRNA transcript level, thus potentially preserving the joint environment.
  • The lack of significant changes in the expression of MMP-2, MMP-9, MMP-13, and SAA suggests that doxycycline’s therapeutic effects may be more targeted, rather than broad-spectrum.
  • Overall, these findings enhance our understanding of the molecular mechanisms underlying doxycycline’s therapeutic effects, and could identify new avenues for equine osteoarthritis treatment strategies in the future.

Cite This Article

APA
Ghasemi S, Sardari K, Mirshokraei P, Hassanpour H. (2018). In vitro study of matrix metalloproteinases 1, 2, 9, 13 and serum amyloid A mRNAs expression in equine fibroblast-like synoviocytes treated with doxycycline. Can J Vet Res, 82(2), 82-88. https://doi.org/10.1155/2015/329418

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 1928-9022
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 82
Issue: 2
Pages: 82-88

Researcher Affiliations

Ghasemi, Samaneh
  • Section of Surgery and Radiology, Department of Clinical Sciences (Ghasemi, Sardari); Center of Excellence in Ruminant Abortion and Neonatal Mortality, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Razavi Khorasan, Iran (Mirshokraei); Department of Basic Sciences, Physiology Division, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Chaharmahal and Bakhtiari, Iran (Hassanpour).
Sardari, Kamran
  • Section of Surgery and Radiology, Department of Clinical Sciences (Ghasemi, Sardari); Center of Excellence in Ruminant Abortion and Neonatal Mortality, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Razavi Khorasan, Iran (Mirshokraei); Department of Basic Sciences, Physiology Division, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Chaharmahal and Bakhtiari, Iran (Hassanpour).
Mirshokraei, Pezhman
  • Section of Surgery and Radiology, Department of Clinical Sciences (Ghasemi, Sardari); Center of Excellence in Ruminant Abortion and Neonatal Mortality, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Razavi Khorasan, Iran (Mirshokraei); Department of Basic Sciences, Physiology Division, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Chaharmahal and Bakhtiari, Iran (Hassanpour).
Hassanpour, Hossein
  • Section of Surgery and Radiology, Department of Clinical Sciences (Ghasemi, Sardari); Center of Excellence in Ruminant Abortion and Neonatal Mortality, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Razavi Khorasan, Iran (Mirshokraei); Department of Basic Sciences, Physiology Division, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Chaharmahal and Bakhtiari, Iran (Hassanpour).

MeSH Terms

  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Cells, Cultured
  • Doxycycline / pharmacology
  • Gene Expression Regulation / drug effects
  • Horses / metabolism
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases / metabolism
  • Real-Time Polymerase Chain Reaction
  • Serum Amyloid A Protein / genetics
  • Serum Amyloid A Protein / metabolism
  • Synovial Fluid / cytology
  • Synoviocytes / metabolism

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