Ensuring agricultural security and preserving the health of wetland ecosystems are crucial concerns facing northeast China. However, the adverse effects of environmental pollution, especially nitrogen (N), caused by prolonged agricultural development on the health of marsh wetlands cannot be systematically recognized. To address this issue, an 18-year trial with four different levels of N application was carried out in a typical area of the Northeast region: 0, 6, 12, and 24 gN·m·a (referred to as CK, N6, N12, and N24, respectively) to investigate changes in wetland ecological functioning. The results showed that long-term N input significantly enhanced soil N availability. High-level of N addition (N24) significantly reduced soil bacterial richness in October, while fungal diversity was significantly higher in June than in October for both control and N6 treatments. The main environmental factors affecting microorganisms in June were TN, NH, and EC, while bacterial and fungal communities were influenced by TN and Leaf Area Index (LAI), respectively, in October. It was found that the AN16S gene was significantly higher in June than in October, indicating that summer is the critical time for N removal in the wetland. N addition significantly reduced the abundance of the NIFH gene and decreased the N fixation potential of the wetland. In June, low and medium levels of N inputs promoted denitrification processes in the wetland and elevated the wetland NO emission potential. The abundance of NARG, NIRK, and NOSZ genes decreased significantly in October compared to June, indicating a decrease in the wetland NO emission potential. Additionally, it was observed that soil methanotrophs were positively affected by NH and TN in October, thereby reducing the wetland CH emission potential. Our research provides a systematic understanding of the impact of agricultural N pollution on marsh wetlands, which can inform strategies to protect wetland health.