Effects of cyclosporin A on human conjunctival fibroblasts.
OBJECTIVE
To evaluate the effects of cyclosporin A (CsA) on cytokine and/or collagen production, cell growth, and apoptosis in conjunctival fibroblast cultures.


METHODS
Fibroblast cultures derived from normal subjects and patients with vernal keratoconjunctivitis and pemphigoid were exposed to different concentrations of CsA for either 24 hours or 30 days. The effects were evaluated by the colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) test to assess cell proliferation, and by the measurement of procollagen I (PIP) and procollagen III (PIIIP) cytokines and total protein in culture medium. CsA-induced apoptosis was assessed by fluorescence-activated cell sorter analysis.


RESULTS
After 24 hours of exposure to doses of CsA of more than 10 microg/mL, cell proliferation and migration were significantly reduced. Cyclosporin A reduced PIP and interleukin 1 (IL-1) production in a dose-dependent manner. Interleukin 6 and IL-8 were increased by 10 microg/mL of CsA, whereas transforming growth factor beta, PIIIP, and total protein were unaffected. Cyclosporin A exposure induced apoptosis in a time- and dose-dependent manner. Long-term exposure to CsA reduced IL-6 but did not modify PIIIP production.


CONCLUSIONS
Exposure to CsA directly modified fibroblast behavior.


CONCLUSIONS
Cyclosporin A ability to accelerate apoptosis in clinically fibrotic tissues may prove to be therapeutic and useful in hyperproliferative conjunctival disorders.
Leonardi
A
A
Ophthalmology and Occular Inflammation Unit, Department of Neuroscience, University of Padova, Italy. mdvol@tin.it
DeFranchis
G
G
Fregona
I A
IA
Violato
D
D
Plebani
M
M
Secchi
A G
AG
D017209:Apoptosis / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003228:Conjunctiva / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D003233:Conjunctivitis, Allergic / Q000378:metabolism / Q000473:pathology; D016572:Cyclosporine / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D010391:Pemphigoid, Bullous / Q000378:metabolism / Q000473:pathology; D011347:Procollagen / Q000096:biosynthesis
D016207:Cytokines; D007166:Immunosuppressive Agents; D011347:Procollagen; D016572:Cyclosporine