AGEs synthesis

AGEs were incubated in vitro following the processes described previously [50]. Briefly, 10g/L of bovine serum albumin (BSA) was co-incubated with 0.5mol/L D-glucose and 1.5mmol/L PMSF in sterile phosphate-buffered saline (PBS) at 37° C for 2 months. The BSA control was incubated similarly but without D-glucose. The solutions were dialyzed in sterile PBS using a dialysis bag (8000~12000 Dalton, 3M, USA), and AGEs and BSA were assessed using a fluorescence spectrometer with an excitation and emission of 370 nm and 440 nm, respectively. The concentrations of AGEs and BSA were detected using a BCA Protein Quantification Kit (Thermo Fisher, USA).