Based on the microstructural investigation of low cycle fatigue under multi-axial non-proportional loading of 316 L stainless steel, the distance within which dislocation moves freely was measured, while its distribution rule was analyzed. The quantitative relation between the macroscopic effective saturated stress and the statistical mean distance within which dislocations moves freely was given. The new definition of non-proportionality of loading path was presented on the basis of the statistical mean distance within which dislocation moves freely, which is considered as entering uniaxial cyclic constitutive equation to describe the stress and strain behavior under multiaxial loading.