Effect of glucagon on proliferation of scleral fibroblast cells of guinea pig in vitro and its mechanism / 中华实验眼科杂志

ABSTRACT

Background The occurrence of experimental myopia may be related to glucagon,and within the range of certain concentration,glucagon may inhibit the development of myopia,but its exact action mechanism is not completely clear.Objective Purpose of this study was to evaluate the effects of glucagon on the proliferation of guinea pig scleral fibroblast cells(GSFCs) and the possible role of glucagon in myopic scleral remodeling.Methods The scleral tissue was obtaincd from the clean blooded guinea pig aged 15 days.GSFCs were cultured and identified with vimentin antibody,cytokeratin antibody and S-100 antibody.0,5,10,50,100,200 μg/L glucagon was added into the different cultured hole for 24 hours respectively,and the growth and proliferation (A490 value) of GSFCs was detected by MTT colorimetric assay.Then the A490 value of GSFCs was assayed in 1 day,2,3,5,7 days under the 50 μg/L glucagon action.Matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2)levels(A450 value)in GSFCs were detected by enzyme-linked immuno sorbent assay (ELISA)72 hours after the cells cultured.Results Passaged GSFCs showed the dendric array at lower density and gyrate array at the higher density with the positive response for vimentin.A490 values of GSFCs were gradually increased with the rise of glucagon concentration(F=32.340,P=0.013).When the glucagon concentration was 10-200 μg/L,the A490Value of GSFCs was higher than that without glucagon group,showing signifitant differences between them(t =5.575,6.627,16.074,12.003,P＜0.05).Under the 50 μg/L glucagon action,A490 values were significantly accented with the time lapse (Ftime =10.610,P =0.024),and the A490 values also were significantly higher than the parallel control groups without glucagon(Fgroup =9.068,P=0.039).MMP-2 level was gradually declined with the enhance of glucagon within range of 5-200 μg/L(F=153.639,P=0.036),but no significant difference was found in TIMP-2 expression(F=24.770,P=3.250).Conclusions Glucagon can promote the proliferation of GSFCs in vitro,and the synthesis of MMP-2shows a concentration-and time-dependent manner.