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)-1βgene 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 800μmol/L. The control group was treated with 0μmol/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-1βgene was measured by qPCR. (2) HTMCs were divided into 3 groups. The control group was treated withserum-free medium and without H2O2 and RSV. The H2O2 group was treated with 300μmol/L H2O2. The H2O2+RSV group was treated with 300μmol/L H2O2 and 25μmol/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 5μg/L TGF-β2. The TGF-β2+RSV group was treated with 5μg/L TGF-β2 and 25μmol/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 800μmol/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-1βgene 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-1βgene 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-1βgene in HTMCs, which may be involved in the development and progression of glaucoma. RSV can inhibit the influence of H2O2 and TGF-β2 in HTMCs and exert a protective effect on glaucoma.