Effect Of IL-1β On MMP-3 Expression In Human Trabecular Meshwork Cells

Purpose:To investigate the effect of Interleukin 1β(IL-1β)on the expression of matrix metalloproteinases-3(MMP-3)in cultured human trabecular cells(HTCs).Methods:(1)In the firt part,the trabecular meshwork tissues from human eyes were primarily cultured.The morphologic features and growing characteristics of the cultured cells were observed by inverted microscope,electron microscope and immunocytochemical medthod.The proliferative curve of the cultured cells was obtained by colorimetric assay with MTT.(2)The 3rd passage cells were incubated with different dosages of IL-1β(0,5,10,25,50 ng/ml,final concentration,diluted by DMEM/F12 without serum)for 24 hours.MMP-3 were determined by RT-PCR and ELISA.Results:(1)The primary cells in cultured were observed from the edge of the cultured tissues about 14 days.The cultured cells were round,oval,fusiform and multiangular.The trabecular cells showed apical villons projections and had a high density of secondary lysosome.The junctions between the trabecular cells observed most frequently were gap junction.(2)Laminin(LN),fibronectin(FN),and neuron specific enolase(NSE)of the cultured HTCs expressed were identified with immunocytochemical medthod.(3) RT-PCR detected that The mean gray scale of MMP-3 /GAPDH expressed by the HTCs treated with 0 ng/ml(control),5 ng/ml,10ng/ml,25 ng/ml,50ng/ml IL-1βfor 24h was0.1037±0.0494,0.3437±0.0764,0.8588±0.0443,1.0079±0.0347,1.3466±0.0309,respectively,The difference between these 5 ng/ml,10ng/ml,25 ng/ml,50ng/ml treated group and that of the control group was statistically significant. (4)ELISA detected that The constration of MMP-3 expressed by the HTCs treated with 0 ng/ml(control),5 ng/ml,10ng/ml,25ng/ml,50ng/ml IL-1βfor 24h was 1.2437±0.3910ng/ml,2.0325±0.2456ng/ml,2.4607±0.3207 ng/ml,2.8532±0.1392 ng/ml,3.5858±0.1141 ng/ml,respectively,The difference between these 5 ng/ml,10ng/ml,25ng/ml,50ng/ml treated group and that of the control group was statistically significant.Conclusion:(1)To master the technology of cell culture.The identification of cultured HTCs must be combined with three aspects:the growing characteristic,morphological features of cultured cells under light and electron microscopes and immunocytochemical medthod.(2)Cultured HTCs expressed MMP-3.(3)In certain extent,IL-1βcan incease expression of MMP- 3 in cultured HTCs.