Abstract:

Objective   To investigate the effect of advanced glycation end products (AGEs) on apoptosis of SHSY5Y cells, and to further explore the relationship between AGEs and the mechanism of Alzheimer disease. Methods   SH-SY5Y cells were treated with different concentrations of AGE-BSA for 48h, or with AGE-BSA (200μg/mL) for different times. Cell apoptosis was detected by flow cytometry (FCM) to determine the best concentration and time of AGE-BSA . SH-SY5Y cells were randomly divided into six groups: the normal control group, the BSA control group, the AGE-BSA group, the AGE-BSA+RAGE antibody group, the AG-BSA+Alpha lipoic acid (ALA) group, and the AGE-BSA+diphenyleneiodonium (DPI) group. Cell apoptosis was detected by FCM and Hoechst staining, the level of ROS was evaluated by the 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) method, and expression of Caspase-12 was analyzed by Western blot. Results   AGE-BSA induced SH-SY5Y cells apoptosis in a time-and concentration-dependent manner. After treatment with 200μg/mL of AGE-BSA for 48 hours, apoptosis of SH-SY5Y cells was significantly increased to (16.8±1.27) % from (2.23±0.08)%(P<0.05). Apoptosis-like cells could be found after Hoechst stained nuclei, the level of ROS and expression of Caspase-12 statistically increased compared with the normal group (P<0.05). Compared with the AGEBSA group, apoptosis of cells, level of ROS and expression of Caspase-12 in the AGEBSA+RAGEAb group, the AGE-BSA + ALA group and the AGEBSA + DPI group were significantly decreased (P＜0.05). Conclusion   AGEs could induce the production of ROS and activation of Caspase-12, which may be involved in apoptosis of SHSY5Y cells induced by AGEs. Blocking the combination of AGEs and its receptor(RAGE) or reducing production of ROS may protect against AGEs-induced SH-SY5Y apoptosis.

Key words: Alzheimer disease; Glycosylation end products, advanced; Reactive oxygen species; Caspase-12; Apoptosis