The objective of this prospective cohort study was to evaluate the gene expression profile of endometrium in dairy cows regarding metritis incidence and parity. Thirty Holstein cows from a group of 90 cows initially enrolled were randomly selected (22 multiparous (MP) and 8 primiparous (PP)) and endometrium biopsies were collected on d 1, 3, and 6 after calving and clinically monitored for metritis. Rectal temperature was measured twice and fever was defined as a temperature ≥39.5°C. A case of metritis was defined as a cow with a red-brown watery foul-smelling uterine discharge or purulent discharge with more than 50% pus and fever on d 1 to 6 postpartum. Twenty-four cows from 30 were selected to analyze the expression of 66 genes measured on the NanoString nCounter Analysis System. The genes selected were related with adhesion, immune system, steroid and prostaglandin biosynthesis regulation, insulin metabolism and transcription factors, and nutrient transporters. The results indicated a different pattern on genes related to immune function by parity. PTX3, involved in antigen presentation, was increased in healthy MP compared with healthy PP whereas inflammatory cytokine TNFα and complement-related protein SERPING1 were upregulated in MP compared with PP (P ≤ 0.05). As expected, metritis incidence affected gene expression pattern related to immune function with a clear increase in expression of antiviral factor MX2 and myosin MYH10 gene, involved in macrophages recruitment, in metritic cows compared with healthy cows (P ≤ 0.05). Differences in uterus involution with metritis were reflected by downregulation of IGF1 (P < 0.10), involved in endometrium remodeling, and a compensatory upregulation of its receptor IGFR1 in metritic cows compared with healthy cows (P ≤ 0.05). A greater expression of receptors of prostaglandins and oxytocin (PGR and OXTR), involved in involution processes, were observed in metritic PP compared with healthy PP (P ≤ 0.05). Overall, these results reflect the effect of metritis in involution and immune response along with the parity influence in post-calving status of the animal.