To explore the effects of different degrees of degradation on soil microbes and soil enzyme activities in alpine meadows, lightly degraded grassland, moderate degraded grassland and severe degraded grassland of alpine grasslands in the Qilian Mountains of the northeastern Qinghai-Tibet Plateau were chosen as the research objects. Characteristic changes of the number of soil microbes (bacteria, fungus, and actinomycetes), soil nitrogen bacteria groups (ammonifier, aerobic nitrogen fixing bacteria, anaerobic nitrogen fixing bacteria and nitrifiers, denitrifying bacteria), soil microbial biomass (soil microbial biomass carbon and soil microbial biomass nitrogen), and soil enzyme activities (sucrose, urease ,phosphatase, and catalase) were determined and analyzed in alpine meadows under different degrees of degradation. The results showed that, for the same soil layer under different degrees of degradation, the number of soil microbes, soil microbial biomass, soil nitrogen bacteria groups, and soil enzyme activities decreased with the increase of degradation degree, and each index was the lowest under severe degradation. The number of actinomycetes, ammoniated bacteria, and denitrifying bacteria in the 10-20 cm soil layer, and the number of bacteria, fungi, actinomycetes, aerobic nitrogen fixing bacteria and denitrifying bacteria in the 20-30 cm soil layer increased in the moderate degraded grassland (p < 0.05). For different soil layers under under the same degrees of degradation, the number of soil microbes, soil nitrogen bacteria groups, soil microbial biomass, and soil enzyme activities decreased with the increase of soil depth. These results provide a new method for assessing the degradation of grasslands, and thus offering important theoretical bases for the restoration of degraded alpine meadows.