Effect Of HA-CD44 Signaling On MMP-2, MMP-9 Expression In Cultured POAG Trabecular Meshwork Cells

Purpose To culture the trabecular cells from patients with primary open-angle glaucoma(POAG) and study their biologic character.To investigate the effect of hyaluronic acid (HA) on the expression of matrix metalloproteinases-2,9(MMPs-2,9) in the trabecular cells,and to study the effect of HA-CD44s signaling on MMP-2,MMP-9 expression.Methods the trabecular tissues from trabeculectomy were cultured in vitro and passaged for 3 times. then identified by immunohistochemistry. to observe its biological characteristics under electronic microscope. Moreover, compare with the normal ones. MMP-2,MMP-9 expressed in cultured trabecular cells were measured by semi-quantitative RT-PCR and gelatin zymography after treated with 0mg∕ml(control),1 mg∕ml,3 mg∕ml,6 mg∕ml HA for 24 huor; anti-CD44,PD9508 was used before HA for 24 hour.Rusults 1. The primarily trabecular cells from patients with POAG were obtained from the edge of tissue after about 10 days . Fibronectin, laminin expression levels in these cells relatively higher, but the ultrastructure of these ones showed the decrease in microvilli, cytolysosome, phagocytic vesicle.2.The relative gray value of MMP-2∕GAPDH of cells treated with0mg∕ml(control),1 mg∕ml,3 mg∕ml,6 mg∕ml HA for 24 huor was 1.11±0.03,1.41±0.04,1.87±0.03,respectively The difference between these 1 mg∕ml,3 mg∕ml,6 mg∕ml HA treated group and that of the control group was statistically significant. The relative gray value of MMP-9∕B-action of cells treated with 0mg∕ml(control),1m∕ml,3mg∕ml,6mg∕ml HA for 24huor,was 1.10±0.05, 1.35±0.06, 2.08+0.04, respectively The difference between these 1 mg∕ml,3 mg∕ml,6 mg∕ml HA treated group and that of the control group was statistically significant.The expressions of MMP-2,9 were decreased when cells were pretreated for 1 hour with anti-CD44 or PD98059. 3.MMP-2 activity of cells treated with 0mg∕ml(control),1 mg∕ml,3 mg∕ml,6mg∕ml HA for 24 huor was assayed by gelatin zymography, 264.50±5.84,353.39±2.56,447.64±5.45, respectively The difference between these 1 mg∕ml,3 mg∕ml,6 mg∕ml HA treated group and that of the control group was statistically significant. MMP-9 activity of cells treated with 0mg∕ml(control),1 mg∕ml,3 mg∕ml,6 mg∕ml HA for 24 huor was assayed by gelatin zymography, 658.51±9.27,895.06±7.15, 1146.75±13.04. respectively The difference between these 1 mg∕ml,3 mg∕ml,6 mg∕ml HA treated group and that of the control group was statistically significant. The expressions of MMP-2,9 were decreased when cells were pretreated for 1 hour with anti-CD44 or PD98059.Conclusion:1.The trabecular meshwork cells from patients with POAG can be successfully cultured in vitro.2.Cultured trabecular meshwork cells espressed MMP-2 and MMP-9.3.HA could increase MMP-2 and MMP-9 espression in cultured trabecular meshwork cells.4. anti-CD44 and PD9508(MEK1 inhibitor) strongly inhibited the HA-dependent MMP-2 and MMP-9 secretion in cultured trabecular meshwork.