The Effects Of Platelet-derived Growth Factor-BB On The Expression Of Matrix Metalloproteinase-2in Cultured Bovine Trabecular Meshwork Cells In Vitro

ObjectiveTo explore complete method of culturing bovine trabecular meshwork cells (BTM cells) in vitro.To investigate the effect and significance of platelet-derived growth factor-BB (PDGF-BB) on matrix metalloproteinase-2(MMP-2) expression in cultured BTM cells in vitro.MethodsCulture the BTM cells in vitro and assessment their origin (expression of NSE,Ⅷ-factor and FN). Count the cells by cytometry and render the cell growth curve. After acquiring the third generation of BTM cells,we preincubate the cells with PDGF-BB (0ng/ml,5.0ng/ml,12.5ng/ml,25.0ng/ml) for24h, to observe the effect of PDGF-BB on the proliferation of BTM cells by SRB and MTT. Preincubated with PDGF-BB (0ng/ml,5.0ng/ml,12.5ng/ml,25.0ng/ml) for2h, the BTM cells were collected to detect the expression of MMP-2by RT-PCR and immunohistochemistry respectively.ResultsTake the third generation BTM cells for characterization. NSE-stained and FN-stained by immunofluorescence was positive. Ⅷ-facter stained by immunofluorescence was negative. The third generation BTM cells get into logarithmic growth phase on the third day and last about three days. Both SRB and MTT demonstrat that PDGF-BB could facilitate the proliferation of BTM cells in vitro in a dose-dependent manner(P<0.05). As concentration of PDGF-BB increased,MMP-2mRNA levels of BTM cells were increased in a dose-dependent manner(P<0.05). Moreover, immunohistochemistry revealed that the MMP-2protein level was distinctly increased in BTM cells stimulated by PDGF-BB(P<0.05).ConclusionThe third day of the third generation BTM cells in vitro were suitable for experiment.PDGF-BB could facilitate the proliferation of BTM cells in vitro, promote the expression level of MMP-2in BTM cells in vitro and may play an important role in decreasing intraocular pressure.