The effect of TGF-beta2 on human trabecular meshwork extracellular proteolytic system

Our study aimed to investigate whether transforming growth factor-beta2 (TGF-beta2), increased in the aqueous humor of eyes with primary open angle glaucoma (POAG), can affect factors responsible for the activity of matrix metalloproteinases (MMPs) in human trabecular cell cultures. With this goal in mind cultures of human trabecular meshwork (hTM) cells derived from 8 donors were treated with TGF-beta2 for 24, 36 and 48 hr. Influence of TGF-beta2 on expression of MMP-2, MMP-9, membrane type 1-MMP (MT1-MMP) and plasminogen activator inhibitor-1 (PAI-1) was examined using RT-PCR, Northern Blot, Western Blot and zymography. The influence of TGF-beta2 treatment on PAI-1 expression was also investigated using immunohistochemistry. It appeared that treatment with TGF-beta2 significantly increased expression of the proform of MMP-2, whereas the active form was not detectable. MMP-9 and MT1-MMP expression were not influenced by TGF-beta2 treatment. There was, however, a significant increase in PAI-1 expression. To investigate whether transformation of the proform of MMP-2 to the active form was inhibited by PAI-1, the influence of treatment with TGF-beta2 and a PAI-1 neutralizing antibody on MMP-2 was investigated using zymogram method. With this treatment protocol the active form of MMP-2 was clearly visible, indicating that TGF-beta2 enhancement of the PAI-1-expression decreases MMP activity. Inhibition of MMP activity through elevated levels of TGF-beta2 might contribute to the increase in ECM in the trabecular meshwork of glaucomatous eyes.