Stationary or mobile microwave radiometers (MRs) can measure atmospheric temperature, relative humidity, and water vapor density profiles with high spatio-temporal resolution, but cannot obtain the vertical variations of wind field. Based on a dataset of brightness temperatures (TBs) measured with a mobile MR over the Three-River-Source Region of the Tibetan Plateau from 18 to 30 July 2021, we develop a direct retrieval method for the wind profile (WP) based on the Long Short-Term Memory (LSTM) network technique, and obtain the reliable dynamic variation characteristics of the WP in the region. Furthermore, the ground-based radiative transfer model for TOVS (RTTOV-gb) was employed to validate the reliability of the TB observation, and we analyzed the impact of weather conditions, altitude, observational mode, and TB diurnal variation on the accuracy of the TB measurement and the retrieval of the WP. Results show that the TB from the mobile observation (MOTB) on clear and cloudy days are close to those of the simulated TB with the RTTOV-gb model, while TB measurements on rainy days are far larger than the modeled TBs. When compared with radiosonde observations, the WPs retrieved with the LSTM algorithm are better than the ERA5 reanalysis data, especially below 350 hPa, where the root mean square errors for both wind speed and wind direction are smaller than those of ERA5. The major factors influencing WP retrieval include the weather conditions, altitude, observational mode, and TB diurnal variation. Under clear-sky and cloudy conditions, the LSTM retrieval method can reproduce the spatio-temporal evolution of wind field and vertical wind shear characteristics. The findings of this study help to improve our understanding of meso-scale atmospheric dynamic structures, characteristics of vertical wind shear, atmospheric boundary layer turbulence, and enhance the assessment and forecasting accuracy of wind energy resources.