Synthesis pattern of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) in human explant organ cultures after treatment with latanoprost and dexamethasone.
OBJECTIVE
To determine changes in production of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the ciliary body, the trabecular meshwork and the retinal pigment epithelium induced by both prostaglandins and corticosteroids.


METHODS
Explant organ cultures were removed by a scleral incision 3 mm posterior to the limbus. Retinal pigment epithelium was grown to confluence. Organ and cell cultures were treated with latanoprost and/or dexamethasone for 72 h. The activity of MMP-2, -3 and -9 was assessed using zymography. The synthesis pattern of MMPs and TIMP-1 and -2 was identified using immunostaining.


RESULTS
Treatment of explant organ cultures with 10 micrograms/ml of latanoprost induced a mean upregulation of MMP-2 by 36%, MMP-3 by 112% and MMP-9 by 156% as seen by zymography. Dexamethasone 500 nm reduced the amounts of secreted MMP-2 by 13%, MMP-3 by 69%. MMP-9 was not detectable in the media of corticosteroid-treated explant organ cultures. The addition of 10 micrograms/ml of latanoprost to dexamethasone-treated cultures increased MMP-2 by 14%, MMP-3 by 43% and MMP-9 by 49%. Using immunohistochemistry we found staining with antibodies against MMP-2, -3, -9 and TIMP-1 and -2 within the ciliary body, and only to a lesser degree in the trabecular meshwork. Latanoprost treatment caused an increase of 29% in MMP-2 (p < 0.0001), 98% in MMP-3 (p < 0.0001) and 108% in MMP-9 (p < 0.0001). Dexamethasone reduced the staining for MMP-2 by 32% (p < 0.0001), for MMP-3 by 33% (p < 0.0001) and for MMP-9 by 83% (p < 0.0001). Almost no change in staining for MMPs was detectable in the trabecular meshwork. Neither latanoprost treatment nor dexamethasone induced significant changes (p < 0.93) in the secretion of TIMPs. In the media of non-treated retinal pigment epithelium (RPE) cells the only MMP detected was MMP-2. RPE cells in culture did not respond to either treatment with a change in their MMP secretion.


CONCLUSIONS
We detected a profound upregulation of both MMP-3 and MMP-9 and a mild induction of MMP-2 through latanoprost in the ciliary body, but not the trabecular meshwork or RPE cells. Corticosteroids, on the other hand, downregulated MMP expression in both tissues. This inhibiting effect of corticosteroids on MMP production was reversed by latanoprost.
el-Shabrawi
Y
Y
Department of Ophthalmology, Karl-Franzens-Universität Graz, Austria. yosuf.elshabrawi@kfunigraz.ac.at
Eckhardt
M
M
Berghold
A
A
Faulborn
J
J
Auboeck
L
L
Mangge
H
H
Ardjomand
N
N
D000368:Aged; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D003907:Dexamethasone / Q000494:pharmacology*; D005123:Eye / Q000187:drug effects* / Q000201:enzymology; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D000077338:Latanoprost; D020782:Matrix Metalloproteinases / Q000096:biosynthesis* / Q000187:drug effects; D008875:Middle Aged; D009924:Organ Culture Techniques; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000201:enzymology; D011461:Prostaglandins F, Synthetic / Q000494:pharmacology*; D019714:Tissue Inhibitor of Metalloproteinases / Q000096:biosynthesis / Q000187:drug effects
D000893:Anti-Inflammatory Agents; D005938:Glucocorticoids; D011461:Prostaglandins F, Synthetic; D019714:Tissue Inhibitor of Metalloproteinases; D000077338:Latanoprost; D003907:Dexamethasone; D020782:Matrix Metalloproteinases