Effect Of Epidermal Growth Factor On Proliferation And Apoptosis In Cultured POAG Trabecular Meshwork Cells And Its Relationship With The High Intraocular Pressure

Objective To demonstrate that if cultured primary open-angle glaucoma trabecullar meshwork cells can secrete epidermal growth factor (EGF)and epidermal growth factor receptor(EGFR),to investigate the effects of EGF on proliferation and apoptosis in POAG trabecullar meshwork cells and the expression of EGFR.Methods the trabecular tissues from trabeculectomy were cultured in vitro and passaged for 3 times. then morphologic feature and growing characteristics of culture cells were observed by inverted microscope,electron microscope and immunocy to chemical method.ELISA was used to detect the expression of EGF and EGFR in POAG trabecullar meshwork cells; The 5th passage cells were incubated with different concentrations of EGF(5 ng/ml,10 ng/ml,20 ng/ml,50 ng/ml,100ng/ml ,final concentration)for 48h respectively, the proliferation and apoptosis effect of EGF on HTCs was detected by Cell Counting Kit-8 assay(CCK-8),and flow cytometry. Using ELISA to detect the expression difference of EGFR before and after the treatment of EGF.Results 1.The primarily trabecular meshwork cells from patients with POAG were obtained from the edge of tissue after about 10 days,the ultrastructure of cells showed microvilli,cytolysosome,phagocytic vesicle,and the expression of fibronectin, 1aminin, neuron specific enolase in these cells were masculine .The cells were identified and confirmed to be POAG trabecular cells.2.The expression of EGF is 1.25ng/ml,and EGFR is 0.89 ng/ml.3.To utilize CCK-8 measure the relative gray value of POAG trabecullar meshwork treat with 0ng/ml(control group),5 ng/ml,10 ng/ml,20 ng/ml,50 ng/ml,100ng/ml EGF were (0.66±0.05),(0.75±0.05),(0.80±0.07),(0.84±0.05),(0.89±0.02),(0.77±0.04),the difference between these 5 ng/ml,10 ng/ml,20 ng/ml,50 ng/ml,100ng/ml EGF treated group and that of the control group was statistically significant. (P<0.05).After the treatment of 5ng/ml,10ng/ml,20ng/ml,50ng/ml,100ng/ml EGF for 48h,the apoptosis rate of trabecular meshwork cells were (14.22±0.18)%,(10.90±0.49)%,(5.98±0.14)%,(5.58±0.21)%,(9.93±0.17)%, showing statistically significant differences with control groups(16.14±0.44)%4.The difference of EGFR expression between 50ng/ml EGF(0.20±0.01)and control group(0.17±0.01)was statistically significant(P<0.05).Conclusion1.POAG trabeeular meshwork cells can be suecessfully cultured in vitro as long as the techllology of cellceultureis known well.2.Trabeeular meshwork cells can secrete EGF and EGFR.3.EGF can promote the proliferation of trabecular meshwork cells, decreases its apoptosis.4.EGF can increase POAG trabecular meshwork cells express EGFR.