Ubiquitin-proteasome system (UPS) plays an important role in neurodegenenetive diseases,such as Alzheimer\'s dis-ease (AD),Parkinson\'s disease (PD),and Huntington\'s disease (HD).The discovery of UPS activators for anti-neurodegenerative dis-eases is becoming increasingly important.In this study,we aimed to identify potential UPS activators using the high-throughput screening method with the high-content fluorescence imaging system and validate the neuroprotective effect in the cell models of AD.At first,stable YFP-CL1 HT22 cells were successfully constructed by transfecting the YFP-CL1 plasmid into HT22 cells,together with G418 screening.The degradation activity of the test compounds via UPS was monitored by detecting the YFP fluorescence in-tensity reflected by the ubiquitin-proteasome degradation signal CL1.By employing the high-content fluorescence imaging system,to-gether with stable YFP-CL1 HT22 cells,the UPS activators were successfully screened from our established TCM library.The repres-entative images were captured and analyzed,and quantification of the YFP fluorescence intensity was performed by flow cytometry.Then,the neuroprotective effect of the UPS activators was investigated in pEGFP-N1-APP (APP),pRK5-EGFP-Tau P301L (Tau P301L),or pRK5-EGFP-Tau (Tau) transiently transfected HT22 cells using fluorescence imaging,flow cytometry,and Western blot.In conclusion,our study established a high-content fluorescence imaging system coupled with stable YFP-CL1 HT22 cells for the high-throughput screening of the UPS activators.Three compounds,namely salvianolic acid A (SAA),salvianolic acid B (SAB),and ella-gic acid (EA),were identified to significantly decrease YFP fluorescence intensity,which suggested that these three compounds are UPS activators.The identified UPS activators were demonstrated to clear AD-related proteins,including APP,Tau,and Tau P301L.Therefore,these findings provide a novel insight into the discovery and development of anti-AD drugs.