Accurate sensing of endogenous microRNAs remains a great challenge for unsatisfactory signal-to-background ratios of traditional techniques. Meanwhile, low sensitivity and insufficient specificity limit the application of most detection methods. Herein, a newly designed GSH-cleavable nanoprobe with the capacity of signal amplification has been proposed for monitoring and imaging of low abundance intracellular miRNA both in a fluorescence assay and in living cells. In this work, endogenous glutathione (GSH) has been introduced to the construction of an miRNA recognizing system for realizing the cyclical process of target. The effective work of GSH makes the circulation of target miRNA possible through spontaneous biocleavage, leading to high detection efficiency and improved sensitivity of nanoprobes. Moreover, minimized false-positive signals and great decrement in environmental disturbance are obtained through ratiometric measurements. Besides, GSH-cleavable nanoprobes provide the enhanced fluorescence resonance energy transfer (FRET) signal between cancer cells and normal cells, which made the practical application in discrimination more reliable.