Activated protein C (APC) regulates matrix metalloproteinase (MMP)-2 production in mouse skin wounds and in human skin keratinocytes. — The Association Specialists
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  3. Activated protein C (APC) regulates matrix metalloproteinase (MMP)-2 production in mouse skin wounds and in human skin keratinocytes.

Activated protein C (APC) regulates matrix metalloproteinase (MMP)-2 production in mouse skin wounds and in human skin keratinocytes. (249)

Sohel Julovi 1 2 , Kelly McKelvey 2 , Nikita Minhas 2 , Aiqun Xue 1 , Ross Smith 1 , Lyn March 3 , Chris Jackson 2
  1. Cancer Surgery Research Group, Department of Surgery, Kolling Institute of Medical Research, University of Sydney at Royal North Shore Hospital, St Leonards, NSW, Australia
  2. Sutton Arthritis Research Laboratory, Kolling Institute of Medical Research, University of Sydney at Royal North Shore Hospital, St Leonards, NSW, Australia
  3. Department of Rheumatology, Royal North Shore Hospital, St Leonards, NSW, Australia

OBJECTIVE: Recently we reported that protease activated receptor (PAR) -2 mediates activated protein C (APC)-induced cutaneous wound healing (Julovi et al, Am J Path, 2011). Matrix metalloproteinases (MMPs) play an important role in tissue remodeling during cutaneous wound healing process.  In this study we investigated the activity of MMP-2 and MMP-9 on murine wound healing and involvement of PAR-2 in the production of MMP-2 induced by APC.

METHODS: Full thickness wounds were made on the dorsum of wild type mice. Mice wounds were treated with APC on days 1, 2 and 3. Human primary keratinocytes were isolated from neonatal foreskin and treated with APC. Wound tissue, culture supernatants and lysates were collected at selected timepoints for zymography, fluorogenic assay (after APMA treatment to measure total MMP), western blotting and immunohistochemical analysis.

 RESULTS:  APC-stimulated wound healing was associated with increased production of MMP-9, and to a lesser extent MMP-2, on day 1 after wounding. In contrast to this early phase of healing, APC almost completely reduced MMP-9 activity while it enhanced MMP-2 activity during the later phase of wound healing on day 6, when APC-treated wounds were >90% healed. To examine the role of PAR-2, human keratinocytes were treated with PAR-2 specific siRNA or blocking antibody to the PAR-2. APC increased production and activation of MMP-2 in culture supernatants, measured by zymography. When PAR-2 was silenced or blocked, MMP-2 production, as measured by fluorogenic assay, was reduced by 44% at 24 hr.

CONCLUSION: APC elevates MMP-9 initially then switches to increase MMP-2 towards the end of cutaneous wound healing. In cultured keratinocytes, APC specifically induces MMP-2 and early evidence suggests that this process requires PAR-2.