Expression of matrix metalloproteinases in wound healing after glaucoma filtration surgery in rabbits.
OBJECTIVE
To investigate the protein and mRNA expressions of matrix metalloproteinases (MMPs), gelatinolytic activity and localization of MMP activity in wounds after glaucoma filtration surgery in rabbits.


METHODS
Sixty eyes of 30 rabbits were removed 1, 3, 7, 14 and 120 days after the surgery and used for this experiment. Protein and mRNA expressions were analyzed by immunohistochemistry and laser capture microdissection/real-time RT-PCR, respectively. The gelatinolytic activity was analyzed by gelatin zymography and the localization was studied using in situ zymography.


RESULTS
By immunohistochemistry, expression of MMP-1, MMP-2, MMP-3, MMP-9 and MT1-MMP was detected in the wounds, most markedly 3 days after the surgery. MMP-positive cells were predominantly macrophages. Expression of MMP-9 and MT1-MMP mRNAs was verified by RT-PCR. Gelatinolytic activities corresponding to proMMP-2 and the active form of MMP-2 were detected in the wounds 3 and 7 days after surgery. In situ zymography localized gelatinolytic activities at the wound site. These activities were almost completely abolished by an MMP inhibitor, indicating that the gelatinolytic activity belongs to metalloproteinases.


CONCLUSIONS
MMPs, particularly MMP-2/MT1-MMP, play important roles in the degradation of the extracellular matrix in the wound healing process after glaucoma filtration surgery and may represent an important target for therapeutic intervention after glaucoma filtration surgery.
Shima
Ichirou
I
Department of Ophthalmology, Kanazawa Medical University, Kahoku-gun, Japan.
Katsuda
Shogo
S
Ueda
Yoshimichi
Y
Takahashi
Nobuo
N
Sasaki
Hiroshi
H
D000818:Animals; D001768:Blister / Q000209:etiology / Q000503:physiopathology; D004210:Dissection / Q000379:methods; D005123:Eye / Q000201:enzymology* / Q000473:pathology; D018463:Filtering Surgery / Q000009:adverse effects; D005780:Gelatin / Q000378:metabolism*; D005901:Glaucoma / Q000601:surgery*; D007150:Immunohistochemistry; D007834:Lasers; D008264:Macrophages / Q000201:enzymology / Q000473:pathology; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D011184:Postoperative Period; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D014945:Wound Healing
D012333:RNA, Messenger; D005780:Gelatin; D020782:Matrix Metalloproteinases; D020778:Matrix Metalloproteinase 2; D053511:Matrix Metalloproteinase 14