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1From the Department of Anatomy II, Friedrich Alexander University, Erlangen, Germany, and 2Department of Ophthalmology, Maximilians-University, Munich, Germany.
PURPOSE. To study whether glaucomatous extracellular matrix (ECM) modifications in the lamina cribrosa might be induced by TGF-ß2, the effect of TGF-ß2 on the expression of collagen types I (Col1
1), III (Col31), and IV (Col42); fibronectin (FN); tissue transglutaminase (TGM2); connective tissue growth factor (CTGF); and thrombospondin (TSP-1) in cultured human optic nerve head (ONH) astrocytes was investigated.
METHODS. Astrocytes were isolated from eyes of five human donors, and cultured monolayers were treated with 1.0 ng/mL TGF-ß2 for 24 and 48 hours. Expression of Col11, Col31, Col42, FN, TGM2, CTGF, and TSP-1 was examined by semiquantitative RT-PCR and Northern and Western blot analyses. The effect of CTGF silencing on the TGF-ß2–modulated expression of these genes was investigated by transfection of CTGF small interfering (si)RNA before TGF-ß2 treatment.
RESULTS. TGF-ß2 treatment upregulated the expression of Col11, Col42, FN, CTGF, TGM2, and TSP-1 mRNA and protein in cultured astrocytes. Inductions ranged between 1.5- and 4-fold. Expression of Col31 remained unaffected. Transfection of 10 nM CTGF siRNA inhibited the TGF-ß2–induced upregulation of CTGF, Col42, Col11, TGM2, and FN, whereas TSP-1 expression was not reduced.
CONCLUSIONS. TGF-ß2 is capable of inducing the expression of ECM and basement membrane components in cultured ONH astrocytes via CTGF and upregulated TSP-1, a protein naturally involved in the activation of latent TGF-ß. Therefore, TGF-ß2 could be a factor in the initiation of the modification of ECM in the glaucomatous ONH. In addition, TSP-1 induction may be a mechanism by which TGF-ß2 amplifies its own activation.
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