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Enclosing plays a crucial role in vegetation and soil quality in grassland. The biomass of green plants, litter, and vertical distributions of soil physical properties and nutrient stocks were evaluated at plot enclosed long term for 38 years inside a fence and a long-term grazing plot outside a fence in a semi-arid grassland of Inner Mongolia. The results showed that dry matter of green plants and litter during the 38-year enclosing treatment was higher than in the grazing treatment (p < 0.01). The soil silt (2–50 μm) in the 38-year enclosing treatment was 5.9% higher than in the grazing treatment (p < 0.01) in 0–10 cm soil, and the fine sand (100–250 μm) was 6.0% lower (p < 0.05). The 38-year enclosing treatment slightly decreased the bulk density and significantly decreased the electrical conductivity in each soil layer (0–100 cm). The 38-year enclosing treatment significantly increased the stocks of soil organic carbon (SOC), available phosphorus (AP), and available potassium (AK) on the surface soil, and obviously decreased the stocks of total nitrogen (TN), total phosphorus (TP), calcium (Ca), magnesium (Mg), sulfur (S), and available nitrogen (AN) in each soil layer (0–100 cm). In conclusion, long-term enclosing improved grassland production, but decreased most nutrient stocks in soil.