Evaluating C3G effects on high glucose-induced redox state

ARPE-19 cells (National Collection of Authenticated Cell Cultures, Shanghai, China) were cultured in Dulbecco’s modified Eagle’s medium (Sigma, USA) with 10% FBS, and the cell culture protocols were referred from Ma, et al. [45 (link)]. To examine the effects of C3G on a high glucose-induced cellular redox state, the cells were cultured in Minimum Essential Medium (MEM) without FBS for 12 h, then pretreated with C3G (10 μM) for 1 h, followed by 30 mM glucose treatment for another 24 h. The cells were used for GSSG/GSH assay, in order to evaluate the cellular redox environment by using the GSH and GSSG Assay Kit (Beyotime Biotechnology, Shanghai, China), respectively, according to the manufacturer’s protocols. Normal glucose (5.5 glucose) groups without C3G pretreatment were used as the control. The high glucose (30 mM glucose) groups without C3G pretreatment were used as the oxidative model.