Sodium nitroprusside(SNP) was used as an exogenous nitric oxide(NO) donor to systematically analyze the regulatory effects of different concentrations(0.0, 0.1, 0.2, 0.3 mmol/L) of SNP solutions on the physiological and biochemical characteristics and shelf life of vine-ripened kiwifruits stored at room temperature after harvest. The results showed that the 0.2 mmol/L SNP treatment had the best preservation effect. It could significantly maintain fruit firmness, reduce weight loss rate and respiration rate, and extend the room temperature shelf life from 8 days for the SNP-free control(CK) group to 14 days, representing a 75% increase. The 0.2 mmol/L SNP treatment delayed the senescence process of vine-ripened kiwifruits through multiple synergistic mechanisms: 1) regulated metabolism and slowing down the consumption of titratable acid(at 8 days of storage, the decrease rate of titratable acid content was slowed by 10.90 percentage points compared to CK), vitamin C(at 8 days of storage, the loss rate of vitamin C was reduced by 18.54 percentage points compared to CK) and dry matter; 2) inhibited the activities of amylase(the peak amylase activity decreased by 18.48% compared to CK), cellulase(peak cellulase activity was only 60.6% of CK) and polygalacturonase, thereby delayed starch degradation and pectin-cellulose network depolymerization, and maintained the firmness of vine-ripened kiwifruit; 3) activated peroxidase(peak peroxidase activity was 1.8 times that of CK), reduced membrane lipid peroxidation levels, and maintained cell membrane structure and function.