Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is an important cofactor in the progress of antioxidant synthesis and biosynthesis, and an abnormal NADPH level has been observed in many viral infection processes. However, efficient tools to monitor NADPH in living cells after viral infection have not been reported. In this work, we present a fluorescent probe, NAFP4, that could detect NADPH ex vivo with a low detection limit of 3.66 nM and image mitochondrial NADPH level changes in living cells. The probe exhibits excellent cell permeability, rapid reactivity, and high selectivity with minimal cytotoxicity. Using NAFP4, we reveal that the NADPH is overproduced in the host cells infected by influenza virus, which was caused by an elevated level of G6PDH during the virus infection. Moreover, there was positive association between the G6PDH level and virus replication. With the proposed probe NAFP4, our study highlights that the virus infection would influence the host metabolism in NADPH production and also suggests that G6PDH is expected to be a promising target for antiviral therapy.
Keywords: G6PDH; NADPH; fluorescent probe; influenza virus; metabolism.