In order to study the effect of intelligent nano-container’s release function in imitation of the condition of Lysosome and Endosome, an acid stimuli-responsive intelligent nano-container using the ketal chain as a stimuli-responsive unit was prepared using Mesoporous Silica Nanoparticles (MSNs) as a carrier, adsorbing fluorescent macromolecular Rhodamine B and encapsulated at the place of MSNs’s mesoporous by β-cyclodextrin with 2-(2-bromo-ethoxy)-2-(2-azido-ethoxy) propane. The functionalized surface of MSNs was demonstrated by Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectrometer (FTIR) and Thermo Gravimetric Analysis (TGA). The releasing performances of Rhodamine B at different pH values were monitored by RF-5201 spectrophotometer and the drug loaded in the intelligent nano-container is 15.23 mg/g which was measured by Ultraviolet-Visible Spectrophotometer (UV-Vis). The intelligent nano-containers constructed by assembly-technology realized “zero release” under neutral condition and remarkable release under acidic condition—suitable release in imitation of the condition of Lysosome and Endosome and this research has extensive application prospects in terms of targeting tumor cells.