HRPE cells were plated into 150 x 15 mm culture plates and incubated at 37[degrees]C in 5% C[O.sub.2].
HRPE cells were plated into 150 x 15 mm culture plates and incubated at 37[degrees]C in 5% C[O.sub.2] until 70-80% confluence after which cells were pre-treated with 25 [micro]M of phloretin, -shogaol and -gingerol individually for 3 h and co-treated with MGO (1 mM) for 96 h.
Briefly, HRPE cells were plated in 150 x 15 mm culture plates and incubated at 37[degrees]C in 5% C[O.sub.2] until 70-80% confluence, then cells were pre-treated with 25 [micro]M phloretin, -shogaol and -gingerol respectively for 3 h and then co-treated with MGO (1 mM) incubated for 96 h.
2, shows that MGO at the low concentrations of 0.1 and 0.5 mM had a negligible effect on the viability of HRPE cells (99 [+ or -] 6% and 98 [+ or -] 4% of cell viability, respectively) following 24 h incubation.
Cytotoxic effects of bioactive compounds on HRPE cells
To evaluate the potential inherent toxicity of the eight specific bioactive compounds (EGCG, ECG, EC, quercetin, -shogaol, phloretin, chlorogenic acid and -gingerol) against HRPE cells, cells were incubated with different concentrations of each bioactive compound for 24 and 96 h after which the cell viability was determined using MTT assay.
Protective effects of the bioactive compounds on MGO-induced damage on HRPE cells
To determine the protective effects of the bioactive compounds on HRPE cells from MGO-induced cell death, cultures were pretreated with bioactive compounds at different concentrations for 3 h respectively prior to co-treatment with MGO at 1 mM for 24 and 96 h period.
5a, shows that bioactive compounds fell into 3 groups for cytotoxicity among which group # 3 was considered to be the most lethal while group # 1 was the least toxic to HRPE cells.
Incubation of HRPE cells with MGO led to an increase in fluorescence intensity due to the formation of AGEs which possess fluorescence properties (Fig.
To postulate the mechanisms by which these bioactive compounds protect HRPE cells against MGO-induced cytotoxicity, western blot analyses were measured on potential therapeutic markers.
HRPE cells treated with MGO alone significantly increased Nrf2 expression in cytoplasm suppressing the defense mechanism (Fig.