Abstract
Purpose: :
To investigate the expression of TNF-α,TNFR1 and NFB in the trabecular meshwork cells of primary open angle glaucoma(POAG) patients . Clarification of the role of TNF-α in the pathophysiology of trabecular meshwork cells, offering new ideas of the pathogenic mechanism, and therapeutic options for POAG.
Methods: :
In vitro cell culture of normal(NTMs) and glaucomatous human trabecular meshwork cells(GTMs).Three samples were cultured from donors eyes respectively. Cells in the third to the fifth generation were collected for the following studies.Immunocytochemistry identification of human trabecular meshwork cells.Immunofluorescence and confocal laser microscopy were used to evaluate the different levels of expression of TNF-α in both NTMs and GTMs.Western blot were applied to test TNF-α,TNFR1 and NFB levels.RT-PCR assays for mRNA quantitation in both NTMs and GTMs.
Results: :
Successful isolation of both NTMs and GTMs on in vitro culture. Immunofluorescence showed significant increased expression of TNF-α in GTMs compared to NTMs.GTMs tested had greater levels of TNF-α,TNFR1 and NFB than NTMs.RT-PCR showed greater mRNA expression of TNF-α in GTMs than the one in NTMs.
Conclusions: :
In vitro culture of both normal and POAG human trabecular meshwork cells expressed TNF-α.The expression of TNF-α,TNFR1 and NFB in GTMs is significantly greater than in NTMs , suggesting TNF-α signaling pathway may involve in the pathogenic mechanisms and progression of POAG.
Keywords: trabecular meshwork • cytokines/chemokines