Abstract:AIM: To investigate the effect of myocilin on the expression of fibronectin(FN)and the adhesion function in trabecular meshwork cells of primary open angle glaucoma(POAG)in vitro.
METHODS: The concentration of recombinant myocilin protein was 0(control group), 0.5, 1, 1.5, 2 and 2.5μg/mL, respectively. The expression of FN in the cells and the cell culture medium were detected by Western blot and ELISA. The effect of myocilin protein on the adhesion of cultured POAG trabecular meshwork cells was detected by CCK-8 method.
RESULTS: Western blot showed that the ratio of FN/β-actin was 34.8±0.6, 33.4±1.0, 28.9±0.8, 21.6±0.9, 15.9±1.1 and 11.9±0.8; that of 0.5μg/mL group was not significantly different compared with that of control group(P=0.092); that of 1.0, 1.5, 2 and 2.5μg/mL group were significantly different compared with that of the control group(F=346.131, P<0.05). The concentration of FN in the cell culture medium was 0.4654±0.0039, 0.4596±0.0032, 0.4216±0.0037, 0.4214±0.0039, 0.4043±0.0039, 0.3806±0.0071μg/mL by ELISA; difference between that of 0.5μg/mL group and that of control group was not statistically significant(P=0.120); the difference between of 1, 1.5, 2, 2.5μg/mL group and the control group were statistically significant(F=176.054, P<0.05). The mean values of the absorbance values of each group were 1.9814±0.1624, 1.8848±0.0267, 1.4895±0.0916, 1.4120±0.1087, 1.3392±0.1391, 1.0310±0.0639 through CCK-8 method; that of 0.5μg/mL group was not significantly different compared with that of control group(P=0.300); the difference between of 1, 1.5, 2, 2.5μg/mL group and the control group were statistically significant(F=177.818, P<0.05).
CONCLUSION: Myocilin protein can inhibit the fibronectin expression in POAG trabecular meshwork cells, showing a dose dependent manner. Myocilin protein can reduce the adhesion of POAG trabecular meshwork cells.
