Protocol implementation for the detection of advanced glycation end products and screening of natural extracts involved in the prevention of diabetic complications
Advanced Glycated End products (AGEs) are involved in the pathogenesis of numerous affections. To improve a protocol for a rapid detection of glycation, in order to screen potential anti-glycation agents, bovine serum albumin (BSA) and methylglyoxal (MG) were incubated at 37 and 50 °C, hourly until 24 h. Electrophoresis and spectrophotometric analysis were used to measure glycated proteins and their aggregations. The anti-AGEs activity screening based on AGEs fluorescence, in 96- well plates, was conducted by incubating BSA and fructose at 50 °C overnight. Quantity of AGEs at 50 °C after 6h of incubation was similar to that at 37 °C after 24h. The screening test showed that four extracts had significant inhibitory effect on BSA glycation (between 40 and 50%). In contrast, some extracts activated this non-enzymatic reaction (between 45 and 60%). The fluorescence determination of BSA (20 mg/ml)-Fructose (0.5 M) system is a rapid and reliable way for anti-glycation phytochemicals screening.
Ramdan Btissam, Ramdan Rajae, Greche Hassane, Bakrim Naima, Nhiri Mohamed. Protocol implementation for the detection of advanced glycation end products and screening of natural extracts involved in the prevention of diabetic complications. J Med Plants Stud 2017;5(3):04-15.