Background and Aim: Uterine lumen contamination with bacteria is ubiquitous in buffalo after parturition. Nearly one-third of these infected animals develop endometritis which leads to reduced fertility. The present study aimed to evaluate the expressions of IFN-γ and NOS2 genes in uterine tissue of buffaloes with endometritis and comparing them with those in healthy animals using RT-qPCR Materials and Methods: Uterine samples were collected from 50 apparently healthy and 50 clinically infected buffaloes. RNA was extracted from the collected buffalo's uteri and cDNA was synthesized from extracted RNA. Quantitative Real Time PCR technique was performed using this synthesized cDNA. Results: Apparent up-regulation of both genes mRNA expression was recorded in endometritis-infected animals with 8.3-folds for IFN-γ and 9.99-folds for NOS2 (P<0.001). Conclusion: The upregulation of IFN-γ and NOS2 expression in the uterine tissue of endometritis-infected buffaloes can be used as a scale for measuring the efficiency of drugs used for endometritis treatment.