The incidence of Inactive ovaries of dairy cows in China is relatively high. There is no complete early warning system for the occurrence of ovarian quiescence in clinical cows. This test provides early warning indicators for clinical prediction of ovary cessation in dairy cows. This experiment selected blood samples of dairy cows from 60 to 90 days postpartum in the inactive ovaries group and control group. Differential proteins were selected on the basis of proteomics, three energy indexes: AST, Glu, NEFA. Four reproductive hormones: E2, P4, FSH, LH, and four differentially expressed proteins: IGFBP-2, AHSG, APO-A4, and RBP-4. Key enzyme activities: ALDOB, LDHB, ITIH3, GPX3, SPAM1, PKM2. The ELISA test kit was used to detect the content and activity of the above markers in the test bovine serum. Through correlation analysis, binary logistic regression modeling and ROC analysis, a single indicator early warning technique for APOA4 and ITIH3 was established. The early warning values were APOA4 > 28.825 μg/L and ITIH3 > 195.07 ng/L. A multi-index early warning system based on potential biomarkers of APOA4 + ITIH3 and APOA4 + ITIH3 + E2 was established. The former had an early warning value of: APOA4 > 19.55 μg/I; ITIH3 > 191.14 ng/L; the latter has an early warning value: APOA4 > 47.56 μg/L, ITIH4 > 187.80 ng/L, E2 < 69.63 ng/L.