Hydrogen peroxide (H 2 O 2 ) release during blood flow is commonly provoked by the cyclic stretch and dynamic shear stress of endothelial cells and is of vital significance for maintaining vascular function. Flexible and stretchable electrochemical sensors show great capability in retrieving mechanical stimulation-induced H 2 O 2 variation; however, cell secretions, especially electroactive constituents' interferences, remain a big concern for sensing accuracy. Herein, we developed a stretchable electrochemiluminescence (ECL) sensor by synthesizing L012-reduced gold nanospheres and decorating them onto a polydimethylsiloxane film-supported gold nanotubes substrate (Au NTs/PDMS) to form dual gold nanostructure-modified meshwork interface. Given the specific reaction between L012 and H 2 O 2 , the as-prepared Au-L012/Au NTs/PDMS exhibited outstanding selectivity toward H 2 O 2 quantification. Through culturing human umbilical vein endothelial cells (HUVECs), real-time monitoring of transient H 2 O 2 release from mechanically sensitive HUVECs in stretching states was realized. This work successfully incorporated the ECL sensing model into in situ cellular sensing, therefore expanding the application mode of the ECL approach for health care and biomedical investigation.