HES1 promotes extracellular matrix protein expression and inhibits proliferation and migration in human trabecular meshwork cells under oxidative stress

Abstract

Glaucoma is the leading cause of irreversible blindness. The most prevalent form of glaucoma is primary open-angle glaucoma (POAG). Oxidative stress is one of the major pathogenic factors of the POAG, and can elicit molecular and functional changes in trabecular meshwork cells, causing increased aqueous humor outflow resistance and elevated intraocular pressure. However, the regulatory mechanisms underlying oxidative stress-induced cell phenotypic changes remain elusive. Herein, we exposed primary human trabecular meshwork cells to the oxidative stress induced by 300 μM H2O2 for 2 h, and found significantly up-regulated expression of extracellular matrix proteins and a transcription factor, hairy and enhancer of split-1 (HES1). The cell functions, including migration and proliferation, were impaired by the oxidative stress. Furthermore, HES1 shRNA abrogated the extracellular matrix protein up-regulation and rescued the functional defects caused by the oxidative stress; conversely, HES1 overexpression resulted in the molecular and functional changes similar to those induced by H2O2. These results suggest that HES1 promotes extracellular matrix protein expression and inhibits proliferative and migratory functions in the trabecular meshwork cells under oxidative stress, thereby providing a novel pathogenic mechanism underlying and a potential therapeutic target to the POAG.

Effect of 3-nitropropionic acid inducing oxidative stress and apoptosis of granulosa cells in geese

“…Our results showed that in goose granulosa cells, treatment with 3-NPA at 5.0 mmol/l down-regulated the mRNA expression of Pcna and p53 , while it up-regulated Caspase 3 mRNA expression. It is plausible that the diminished proliferative and augmented apoptosis caused by the transient oxidative stress induced by 3-NPA may contribute to the inhibition of granulosa cell viability in geese [ 30 ]. The anti-apoptotic protein Bcl-2 inhibits cytochrome c release and Caspase 3 activation and thus functions as an antioxidant to prevent apoptosis [ 31 ].…”

Lentivirus-mediated IL-10-expressing Bone Marrow Mesenchymal Stem Cells promote corneal allograft survival via upregulating lncRNA 003946 in a rat model of corneal allograft rejection

“…All the lentivirus-involving experiments, including packaging, concentration, tittering, and injections, were performed in a dedicated Biosafety Level 2 microbiology laboratory, and all the experimental procedures adhered to the health and safety guidelines stated by BIOSAFETY LEVEL 2 LABORATORY PRACTICES from NIH. The packaging of lentivirus was performed as described previously 43 , 44 . Briefly, 293T cells were seeded on the 6-well plates (Corning, Corning, NY, USA) coated by rat tail collagen (Sigma-Aldrich, St. Louis, MO, USA) at a density of 8 × 10 5 cells / well.…”

“…Cells were transduced as previously described 43 , 44 . Briefly, the titers of Lenti-IL-10 and Lenti-RFP were equally adjusted.…”

“…compound (Sakura Finetek, Torrance, CA, USA), frozen in liquid nitrogen, and sectioned into 5 μm thickness. Immunofluorescence was performed as previously described 43 , 44 , 47 . Briefly, the sections were post-fixed in 4% PFA, washed with PBS, permealized with 0.03% sodium dodecyl sulfate, and blocked with 10% goat serum.…”

Oxidative stress and antioxidants in the trabecular meshwork

“…HES1 regulates the development of cells in the nervous and digestive systems by functioning downstream of the Notch signaling pathway (Kageyama, Ohtsuka & Kobayashi, 2007). Xu et al found that HES1 promotes ECM expression and inhibits TMC proliferation and migration under oxidative stress (Xu et al, 2017). More importantly, HES1 short hairpin RNA (shRNA) has been shown to attenuate ECM protein upregulation and functional defects caused by oxidative stress.…”