Abstract: Abstract: Objective To investigate hydrogen peroxide (H2O2) and transforming growth factor-β2 (TGF-β2) induced fibronectin (FN), collagen 1 (COL1), nuclear factor (NF) -κB P65 proteins and interlukin (IL) -1β gene expression in human trabecular meshwork cells (HTMCs), and the interventional mechanism of resveratrol (RSV). Methods (1) HTMCs with 70 to 80% confluency were divided into 5 groups. The experimental groups were treated with serum-free medium and with H2O2 at concentrations of 150, 300, 450 and 800 μmol/L. The control group was treated with 0 μmol/L H2O2. The protein levels of FN, COL1, NF- κB P65 and NF- κB P65 phosphorylation (P- NF- κB P65) were measured by Western blot assay. The expression of IL-1β gene was measured by qPCR. (2) HTMCs were divided into 3 groups. The control group was treated with serum-free medium and without H2O2 and RSV. The H2O2 group was treated with 300 μmol/L H2O2. The H2O2 +RSV group was treated with 300 μmol/L H2O2 and 25 μmol/L resveratrol (RSV). The expressions of proteins and genes mentioned above were detected in three groups. NF-κB P65 nuclear translocation was assessed by immunofluorescence technique. (3) HTMCs were divided into 3 groups. The control group was treated with serum-free medium and without TGF-β2 and RSV. The TGF- β2 group was treated with 5 μg/L TGF-β2. The TGF-β2 +RSV group was treated with 5 μg/L TGF-β2 and 25 μmol/L RSV. The expressions of proteins and genes mentioned above were detected in three groups. Results (1) Compared with control group, the protein levels of FN and P-NF-κB P65 were significantly increased in 150, 300, 450 and 800 μmol/L groups， the expression levels of COL1 protein and IL-1β gene were significantly increased in 300, 450 and 800 μmol/L groups (P < 0.05). There were no statistical significances between other indicators. (2) The expression levels of FN, COL1, P-NF-κB P65 proteins and IL-1β gene were significantly higher in H2O2 group than those in control group, and which were significantly lower in H2O2 +RSV group than those in H2O2 group. Compared with control group, only the expression of IL-1β gene was decreased in H2O2 +RSV group (P < 0.05). NF- κB P65 was only expressed in cytoplasm in control group, while it was expressed in both cytoplasm and nucleus in H2O2 group. Compared with H2O2 group, NF-κB P65 was mainly expressed in cytoplasm. (3) Compared with control group, the expressions of FN, COL1, P-NF-κB P65 proteins and IL-1β gene were significantly increased in TGF-β2 group (P < 0.05). Compared with TGF-β2 group, the indicators mentioned above were significantly decreased in TGF-β2+RSV group (P < 0.05). Conclusion H2O2 and TGF-β2 can upregulate the expression of FN, COL1, P-NF-κB P65 proteins and IL-1β gene in HTMCs, which may be involved in the development and progression of glaucoma. RSV can inhibit the influence of H O and TGF-β in HTMCs and exert a protective effect on glaucoma.

Key words:  glaucoma, trabecular meshwork, hydrogen peroxide, transforming growth factor beta2, extracellular matrix, NF-kappa B, interleukin-1beta, protein fiber connection； collagen 1； Resveratrol