Quantitative microstructural evolution and the corresponding microhardness of electrodeposited nanostructured nickel sheet during cold rolling deformation are investigated by x-ray diffraction, transmission electron microscopy and Vicker’s microhardness testing. Particularly, to investigate the effect of stress states on deformation behavior, two series of gradient nanostructured nickel with symmetric structures and the homogeneous counterparts with three levels of grain size are compared based on macro-statistical data. In such hierarchical sandwich-like gradient samples, the layers with larger grain size, as the soft phase, indeed sustain more deformation. Deformation-induced grain rotation changes are observed in the center layers with a relatively larger grain size, accompanied by an obvious decrease in microhardness. According to the quantitative microstructural parameters including the grain size, dislocation density and stacking fault probability before and after deformation, evaluation based on Hall-Petch and Bailey-Hirsch relationships indicates the transition from strain hardening to softening can be attributed to grain orientation change.