RESUMO
Previous studies have shown that sirtuin 1 (SIRT1) reduces the production of neuronal amyloid beta (Aß) and inhibits the inflammatory response of glial cells, thereby generating a neuroprotective effect against Aß neurotoxicity in animal models of Alzheimer's disease. However, the protective effect of SIRT1 on astrocytes is still under investigation. This study established a time point model for the clearance of Aß in primary astrocytes. Results showed that 12 hours of culture was sufficient for endocytosis of oligomeric Aß, and 36 hours sufficient for effective degradation. Immunofluorescence demonstrated that Aß degradation in primary astrocytes relies on lysosome function. Enzymatic agonists or SIRT1 inhibitors were used to stimulate cells over a concentration gradient. Aß was co-cultured for 36 hours in medium. Western blot assay results under different conditions revealed that SIRT1 relies on its deacetylase activity to promote intracellular Aß degradation. The experiment further screened SIRT1 using quantitative proteomics to investigate downstream, differentially expressed proteins in the Aß degradation pathway and selected the ones related to enzyme activity of SIRT1. Most of the differentially expressed proteins detected are close to the primary astrocyte lysosomal pathway. Immunofluorescence staining demonstrated that SIRT1 relies on its deacetylase activity to upregulate lysosome number in primary astrocytes. Taken together, these findings confirm that SIRT1 relies on its deacetylase activity to upregulate lysosome number, thereby facilitating oligomeric Aß degradation in primary astrocytes.