Effect Of Fibronectin On Proliferation,adherence And Immigration As Well As Integrinα5β1,α4β1 Expression In Cultured POAG Human Trabecular Cells

Purpose To culture the trabecular cells from patients with primary open-angle glacoma(POAG) and study their biologic character. To investigate the effect of fibronectin (FN) on the expression of proliferation, adherence and Immigration as well as integrinα5β1,α4β1 in the trabecular cells, and to study the effect of fibronectin signaling on integrinα5β1,α4β1 expression.Methods The trabecular tisses from trabeclectomy were cultured in vitro and passaged for 3 times. then identified by Immunocytochemical stain to observe its biological characteristics under electronic microscope. Moreover, Effect of fibronectin on proliferation, adherence and Immigration in cultured human trabecular cells were measured by Cell Conting Kit-8 and Transwell ventricle after treated with 0μg/ml (control),5μg/ml,10μg/ml,20μg/ml,40μg/ml,100μg/ml FN for 24 hour and expression of fibronectin signaling on integrinα5β1,α4β1 expression.Results 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 masculine, but the ultrastructure of these ones showed microvilli, cytolysosome, phagocytic vesicle.2. To utilize Cell Conting Kit-8 and Transwell ventricle measure the relative gray value of POAG trabecular meshwork cells proliferation treated with5μg/ml,10μg/ml,20μg/ml,40μg/ml, 100μg/ml FN for 24 hour was 0.45±0.016,0.52±0.015 0.65±0.011,0.78±0.012,1.15±0.014 in experiment, compare with0.37±0.018 control respectively P<0.05 The difference adherence between these 0.38±0.021, 0.43±0.015,0.52±0.021,0.78±0.025,0.85±0.014,1.19±0.01treated group and that0.38±0.021 of the control group was statistically significant. Difference Immigration between these 0.59±0.005,0.62±0.006,0.68±0.002, 0.78±0.003,0.89±0.001 the control 0.239±0.011group was statistically significant(P<0.05).The identified by Immunocytochemical stain to measure the relative gray value of POAG trabecular meshwork cells treated with5μg/ml,10μg/ml,20μg/ml,40μg/ml,100μg/ml FN for 24 hour was the expressions of ntegrinα5β1 was 0.242±0.015,0.254±0.018,0.261±0.005 0.267±0.014,0.365±0.019, compare with 0.239±0.011the control group was statistically significant (P<0.05). Masculine areas were repectively0.1428±0.013 0.1946±0.018 0.2219±0.016 0.2679±0.011 0.17993±0.019, compare with 0.1252±0.012(control) group was statistically significant(P<0.05).The expressions of integrinα4β1 was 0.232±0.005,0.245±0.003, 0.263±0.005,0.276±0.004,0.385±0.019 compare with 0.231±0.001 the control group was statistically significant(P<0.05). Masculine areas were repectively 0.1218±0.013 0.1346±0.016 0.1419±0.012 0.1679±0.011 0.21993±0.019, compare with 0.120±0.012 (control) group was statistically significant(P<0.05).Conclusion:1. The trabecular meshwork cells from patients with POAG can be successfully cultured in vitro.2. Cultured trabecular meshwork cells expressed integrinα5β1 and integrinα4β1.3. In invariably circumscription, effect of fibronectin could increase on proliferation, adherence and Immigration about dose dependent in cultured human trabecular cells4. In invariably circumscription, FN could increase integrinα5β1 and integrinα4β1 expression about dose dependent in cultured trabecular meshwork.