Modulation of HTM Cell Responses

HTM cells were purchased from Meisen Chinese Tissue Culture Collections (Zhejiang, China). The cells were cultured in Dulbecco's Modified Eagle Medium (DMEM)/Nutrient Mixture F-12 medium (L310KJ; BasalMedia, Shanghai, China) containing 15% fetal bovine serum and supplemented with 1% penicillin–streptomycin (GA3502; GeneView, Beijing, China). Cultures were maintained at 37°C with 5% CO₂. HTM cells were verified by detecting the expression of myocilin after treatment with 100-nM dexamethasone (HY-14648; MedChemExpress, Monmouth Junction, NJ) for 5 days.²⁹ (link)^,30 (link) The cells were treated as follows: for the control (CON), NR, H₂O₂, and TGF-β2 groups, HTM cells were incubated with culture medium, 1-mM NR (S31692; Shanghai Yuanye Bio-Technology, Shanghai, China), 200-µM H₂O₂ (E882, AMRESCO, Solon, OH), and 10-ng/mL TGF-β2 (Z03429-50; GenScript, Jiangsu, China), respectively, for 48 hours. For the NR+H₂O₂ group, cells were pretreated with 1-mM NR for 24 hours and then incubated with 200-µM H₂O₂ for another 24 hours. For the TGF-β2+NR group, cells were treated with 10-ng/mL TGF-β2 and 1-mM NR for 48 hours simultaneously. H₂O₂ and NR were diluted in the medium to reach the final concentrations.