An Experimental Investigation Of Diclofenac Sodium And Ketorolac Tromethamine On Influence Of Rabbit Corneal Stromal Cells And Fibronectin

Objective To discuss the effect of diclofenac sodium and Ketorolac tromethamine on inhibiting of rabbit corneal stromal cells and influence of fibronectin. Explore it's mechanism. Methods The primary culture or subculture of RCSCs were exposed to different concentrations of Diclofenac sodium,Ketorolac tromethamine and Fluorometholone in experimental group and to culture media in control group. The inhibitive characteristics to the cells' proliferation were measured with methyl thiazolyl tetrazolium(MTT) assay. The morphological changes of cells were observed with light microscope and transmission electron microscope. The changes of cells and the apoptosis rate were observed with flow cytometer(FCM). The changes of the apoptosis indexes were measured by immunocytochemistry. An enzyme-linked immunohistochemistry assay(ELISA) was used to determine the content of the fibronectin(FN)in the supemate. Results Diclofenac sodium , Ketorolac tromethamine and Fluorometholone can inhibit proliferation of RCSCs, while Diclofenac sodium showed strongest antiproliferative effect. And Diclofenac sodium showed marked dosage dependence and time-effect, the two others also had dosage dependence and not obviously time-effect. FCM result of Diclofenac sodium and Ketorolac tromethamine showed that the G0/G1 phase cells obviously increased and the apoptosis cusp and rate increased. Immunocytochemistry showed Bcl-2 and Bax increased while Bax increased even more in Diclofenac sodium and Ketorolac tromethamine, so the ratio of Bax/Bcl-2 increased. ELISA demonstrated increase in supemate FN content following the increasing concentrations of Diclofenac sodium. And the FN content in supemate increased in the 50μg/ml of Ketorolac tromethamine, while the FN content in supemate decreased following the increasing concentrations in the others group. Conclusion 1,Diclofenac sodium,Ketorolac tromethamine and Fluorometholone has inhibitive effect on RCSCs and Diclofenac sodium showed stronger effect. 2,Diclofenac sodium and Ketorolac tromethamine may exert theirs antiproliferative effect by inducing apoptosis and ceasing cells cycles. 3,Diclofenac sodium and Ketorolac tromethamine had been obviously different on influence of extracellular matrix(ECM). 4,Ketorolac tromethamine may be similar with steroid on the mechanism of preventing refractive regression.