2, AGE-BSA increased intracellular ROS levels, and pretreatment with OSSC extracts significantly attenuated this increase, with 100ng/ml OSSC extracts reducing AGE-BSA-stimulated intracellular ROS levels to ~80% of those in cells treated with AGE-BSA alone.
To test whether OSSC extract could prevent AGE/RAGE binding activity, purified hRAGE protein was incubated with HRP-labelled AGE-BSA and various concentrations of OSSC extract in vitro.
4, AGE-BSA induced NF-[kappa]B nuclear translocation in GMCs, and OSSC extract induced on NF-[kappa]B translocation from the nucleus to the cytosol in AGE-BSA-induced GMCs.
5A-D shows representative immunoblots of phosphorylated MAPK (p38 and ERK1/2) and I[kappa]B from cells cultured in AGE-BSA, with or without OSSC extract pretreatment.
2, fluorescence excitation and emission spectra of hydrolyzed BSA and peptide-derived AGE calibrator (obtained by degradation of AGE-BSA with proteinase K) are presented.
Calibration was carried out with a single calibrator of AGE-BSA (1 mmol/L AGEBSA = 12 [A.
The analytical signals for hydrolized calibrators of AGE-BSA ([S.
As no AGE standard is available, external calibration was carried out with an AGE-peptide calibrator derived from an AGE-BSA calibrator (50 g/L) by hydrolysis with proteinase K.