The current designs of ratiometric fluorescence sensing strategies primarily make use of the opposite responses of analyte target to two fluorescence probes, often overlooking their potential capabilities for signal amplification. Herein, a novel ratiometric fluorescence sensor was constructed for sensitive detection of glutathione (GSH) by combining CeO2/CuO nanozymes with poly[2,7-(9,9′-dioctylfluorene)-alt-4,7-bis(thiophen-2-yl) benzo-2,1,3-thiadiazole] (PFO-DBT)-based polymer dots (Pdots). The CeO2/CuO nanozymes with high oxidase-like activity were prepared via a deep eutectic solvent (DES) strategy and used for catalytic oxidization of o-phenylenediamine to produce fluorescent 2,3-diaminophenazine (DAP), which concurrently quenched the fluorescence of Pdots through photoinduced electron transfer (PET), thus amplifying the ratiometric signal. Upon addition of target GSH, the oxidase-like activity of CeO2/CuO were inhibited due to its blocking to metal active sites and scavenging superoxide anion (O2‧−), which responsively decreased the amount of produced DAP, and thus increased the fluorescence of Pdots. The designed CeO2/CuO-OPD-Pdots system showed a linear ratiometric response to GSH concentration within the range of 2 μM to 20 μM, with a limit of detection (LOD) determined as 1.24 μM. The proposed method could be used for selective detection of GSH in serum samples, and thus possessed potential application in clinic analysis.