Effect Of Myocilin On Fibronectin Expression And Cell Adhesion And Migration In Cultured Primary Open-angle Glaucoma Trabecular Meshwork Cells

Purpose To investigate the effect of myocilin on fibronectin expressionand cell attachment and migration on the basis of culturing the trabecular meshworkcells from primary open-angle glaucoma eye，to study the relationship betweenmyocilin and POAG.Methods The trabecular tissues from trabeculectomy were cultured in vitroand passaged for3times.The third passage cells were incubated with different finalconcentrations of recombinant myocilin(0ug/ml(controlgroup)、0.5ug/ml、1ug/ml、1.5ug/m、2ug/ml、2.5ug/ml)，which were diluted by DMEM/F12without serumfor24hours. Western blot and Elisa were used to detect the fibronectin expressionin cultured trabecular meshwork cells from POAG.CCK-8assay and transwellchamber were used to detect adhesion and migration ability of thetrabecular meshwork cells from POAG.Results1、The primarily trabecular meshwork cells in vitro from POAG throughcontinuous cell culture inverted microscope and immunohistochemistry wereidentified as purified trabecular meshwork cells;2、western bolt result display:33.4±1.0、28.9±0.8、21.6±0.9、15.9±1.1、11.9±0.8.The0.5ug/ml myocilinexperimental group compared with the control group, the difference was notstatistically significan（tP=0.09，P>0.05）. The1ug/ml、1.5ug/ml、2ug/ml、2.5ug/mlmyocilin experimental group compared with the control group and within these threeexperimental groups the difference were statistically significant(P=0.00，P<0.05).3、The fibronectin concentration of trabecular cells were0.4654±0.0039ug/ml、0.4596±0.0032ug/ml、0.4216±0.0037ug/ml、0.4214±0.0039ug/ml、0.4043±0.0039ug/ml、0.3806±0.0071ug/ml by Elisa assay. The0.5ug/mlmyocilin experimental group compared with the control group, the difference was not statistically significan（tP=0.12，P>0.05）. The1ug/ml、1.5ug/ml、2ug/ml、2.5ug/mlmyocilin experimental group compared with the control group and within these threeexperimental groups the difference were statistically significant(P=0.00，P<0.05).4、Our results showed that there were more migrated cells in controlgroup than in experimental group(control group were16.40±4.0373，experimental group were8.60±2.0736), and the difference between themwas statistically significant(P=0.005，P<0.05）.5、The OD value of trabecularmeshwork cells were1.9814±0.1624、1.8848±0.0267、1.4895±0.0916、1.4120±0.1087、1.3392±0.1391、1.0310±0.0639by CCK-8assay. The0.5ug/mlmyocilin experimental group compared with the control group, the difference was notstatistically significant（P=0.30，P>0.05）. And the1ug/ml、1.5ug/ml、2ug/ml、2.5ug/ml myocilin experimental group compared with the control group andwithin these three experimental groups the difference were statistically significant（P=0.00，P<0.05）.Conclusion1、Myocilin protein could reduce the fibronectin expression intrabecular meshwork cells, and showed a dose dependent manner.2、The migrationand adhesion ability of trabecular meshwork cells were inhibited by myocilin.