Background: Root canal treatment aims to eliminate bacterial presence and debris from within the root canal system. Material and Methods: This study utilized sixty mandibular premolars with single roots, which underwent decoronation and root canal shaping. Post-sterilization, the specimens were inoculated with Enterococcus faecalis and incubated at 37°C for 48 hours. Subsequently, the specimens were segregated into six groups, differentiated by their irrigation solutions: 2.5% sodium hypochlorite, 5.25% sodium hypochlorite, a solution derived from Moringa oleifera leaves, a mixture of 2.5% sodium hypochlorite with M. oleifera leaf solution, a mixture of 5.25% sodium hypochlorite with M. oleifera leaf solution, and a control group treated with saline. Microbial sampling occurred pre-irrigation, post-irrigation, and seven days post-irrigation to assess bacterial reduction via colony-forming unit (CFU) counts. Results: ANOVA, or two-way repeated measures and Tukey's HSD revealed a notable decline in CFU/ml across all experimental groups after irrigation. Noteworthy is the significant microbial reduction observed in groups irrigated with sodium hypochlorite solutions combined with M. oleifera leaf extract compared to those irrigated with sodium hypochlorite alone. Conclusion: The findings suggest the potential of Moringa oleifera leaf extract as a viable endodontic irrigant, offering a significant antibacterial effect, particularly when used in conjunction with traditional sodium hypochlorite solutions.