Wheat is the third most producing crop in China after maize and rice. In order to enhance the nitrogen use efficiency (NUE) and grain yield of winter wheat, a two-year field experiment was conducted to investigate the effect of different nitrogen ratios and doses at various development stages of winter wheat (Triticum aestivum L.). A total of five N doses (0, N75, N150, N225, and N300 kg ha−1) as main plots and two N ratios were applied in split doses (50%:50% and 60%:40%, referring to 50% at sowing time and 50% at jointing stage, 50% at sowing time + 50% at flowering stage, 50% at sowing time + 50% at grain filling stage, and 60% + 40% N ratio applied as a 60% at sowing time and 40% at jointing stage, 60% at sowing time and 40% at flowering stage, and 60% at sowing time and 40% at grain filling stage in subplots). The results of this study revealed that a nitrogen dose of 225 kg ha−1 significantly augmented the plant height by 27% and above ground biomass (ABG) by 24% at the grain filling stage, and the leaf area was enhanced by 149% at the flowering stage under 60 + 40% ratios. Furthermore, the N225 kg ha−1 significantly prompted the photosynthetic rate by 47% at the jointing and flowering stages followed by grain filling stage compared to the control. The correlation analysis exhibited the positive relationship between nitrogen uptake and nitrogen content, chlorophyll, and dry biomass, revealing that NUE enhanced and ultimately increased the winter wheat yield. In conclusion, our results depicted that optimizing the nitrogen dose (N225 kg/ha−1) with a 60% + 40% ratio at jointing stage increased the grain yield and nitrogen utilization rate.