Nitrous oxide (N2O) is a potent and long-lived greenhouse gas that contributes to global warming with a global warming potential (GWP) 298 times that of carbon dioxide (CO2). In this paper, we analyzed the trend of N2O concentration in vertical layers of the stratosphere from 2005 to 2020 using the N2O observed from the Microwave Limb Sounder (MLS) that is on board the Aura satellite. We found that the local N2O concentration showed a downward trend in the lower stratosphere but rose or fluctuated in the upper stratosphere. The reduction reached −5 ppb/yr at pressure levels of 31.62 hPa and 68.13 hPa, with a confidence level of over 90%. The growth was around 1–2 ppb/yr in the upper stratosphere. In addition, a concentration anomaly was observed in the tropical stratosphere in 2013. After the appearance of this anomaly, the N2O concentration in the middle and lower layers of the tropical stratosphere was lower than before 2013. We speculated that the enhancement of the Brewer–Dobson circulation (BDC) upwelling before and after stratospheric sudden warming (SSW) is the main reason for the abnormal concentration distribution in 2013. Stratospheric N2O has changed significantly in the past 16 years with the mutual coupling effect of BDC and SSW and such changes can have further impact on the chemical equilibrium and radiation balance in the stratosphere, as well as on the persistent climate-warming trend.