[Objective] Human disturbance is a critical factor causing grassland degradation, therefore, investigation of its impact against grassland soil quality and healthiness will provide a scientific basis for ecological restoration, management strategy formulation, and sustainable utilization of grassland resources. [Methods] This study was conducted to measure soil physical and chemical properties and plant biomass under four disturbance regimes as enclosure, grazing, reclamation, and tourism in Xilamuren Grassland of Inner Mongolia. The comprehensive healthiness of soil and Grassland were evaluated using grey functional analysis and fuzzy comprehensive evaluation methods. [Results] ① The highest percentages of clay (15.45%) and silt (20.13%) were found in enclosure zones whereas the lowest (7.21% and 10.02%) value were detected in tourism zones. The highest soil bulk density (1.61g/cm3) was also found in tourism zones. No significant differences were detected for soil moisture and saturated water-holding capacity within soil layers of sampling sites. ② The highest soil organic matter (12.98 g/kg) and total nitrogen (0.73 g/kg) were observed in enclosure zones while the lowest (8.23 g/kg and 0.46 g/kg) values were found in tourism zones. Total phosphorus and potassium levels among different human disturbance were ranked as reclamation > enclosure > grazing > tourism. 9.08 mg/kg more available phosphorus in reclaimed areas was higher than tourism zones while the content of available potassium (268 mg/kg) in enclosure zones was significantly higher than other sites. ③ Total plant biomass of sampling site was ranked as enclosure (342.67g/㎡) > reclamation (155.29g/㎡） > grazing (112.93g/㎡) > tourism (43.51g/㎡). ④Soil quality and comprehensive grassland healthiness was comprehensively ranked as the order of enclosure > reclamation > grazing > tourism. [Conclusion] Significant differences in soil physicochemical properties and plant biomass were found under different human disturbance regimes. Enclosure zones demonstrated the best soil quality and grassland healthiness while tourism zones showed the worst condition. Both reclamation and grazing negatively impacted soil quality and grassland healthiness. These findings will provide a scientific foundation for grassland ecological restoration, management planning, and sustainable development strategies.