Diabetes is a metabolic syndrome associated with hyperglycemia, hypertension, atherosclerosis, and endothelial dysfunction. Applying the mechanical stretch on cells to simulate blood circulation while monitoring the cell glucose metabolism in a high-glucose environment is important for better comprehension of the underlying mechanisms of atherosclerosis caused by diabetes. Herein, we developed a facile strategy integrating zeolitic imidazolate framework-8-encapsulated glucose oxidase (GOx@ZIF-8) and an gold (Au) stretchable electrode (Au SE) to construct a flexible and stretchable glucose sensor (GOx@ZIF-8/Au SE) for investigating the glucose metabolism mechanism of stretched endothelial cells in hyperglycemia. The encapsulation of GOx with ZIF-8 prevents the aggregation and detachment of GOx from the sensing interface and endows the biosensor with high stability. Additionally, the Au SE with inherent stretchability can act as an integrated platform for mechanical stimulation as well as for transient signal sensing during the mechanotransduction process. Moreover, this flexible and stretchable glucose sensor is successfully used for monitoring the glucose metabolism of mechanically stimulated cells in hyperglycemia, and it was found for the first time that the glucose utilization ability of cells under static conditions is higher than that in the stretched state. This facile and straightforward method paves a promising route for designing a stable enzyme-based stretchable biosensor for detecting the underlying mechanisms of atherosclerosis caused by diabetes.