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Recent studies have confirmed that sulforaphane (SFN) can activate multiple pathways, and promote the expression of the antioxidants in cells.Thioredoxin (Trx) plays an important role in maintaining the intracellular redox in the steady state.
This study was to investigate the effect and mechanism of SFN on Trx expression in bovine trabecular meshwork cells (BTMCs) cultured in vitro.
BTMCs were cultured in vitro and identified by morphological evaluation.The third generation of BTMCs were cultured in the medium with 0, 10, 20 and 30 μmol/L SFN for 30 minutes.Real-time PCR was applied to measure the expression of Trx mRNA in BTMCs.The BTMCs were randomly divided into normal control group, LY294002 group, U0126 group, SFN group, LY294002+ SFN group and U0126+ SFN group.The expressions of Nrf2 protein and Trx protein in each group were measured by Western blot.
The BTMCs was successfully cultured in vitro.The expressions of Trx mRNA were significantly different among the different concentrationss of SFN treatment (F=88.090, P<0.01). The expressions of Trx protein and Nrf2 protein in the LY294002+ SFN group, U0126+ SFN group and SFN group were significantly higher than those in the normal control group (all at P<0.01). The expressions of Trx protein and Nrf2 protein in the LY294002+ SFN group and U0126+ SFN group were significantly higher than those in the SFN group (all at P<0.01).
SFN can activate Nrf2 by phosphatidylinositol-3-kinase(PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases 1/2 (ERK1/2) signaling pathways, which can increase the expression level of Trx in BTMCs cultured in vitro.