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Abstract Background This study aimed to investigate the antiparasitic and anti-inflammatory potential of pumpkin seed oil in mice infected with Trichinella spiralis by demonstrating its impact on MMP-9 expression and pathogenesis during the intestinal and muscular phases. Results In this study, 100 mice were divided into five groups: an infected group, a pumpkin seed oil-treated group (1.5 mg/kg BW, administered three times per week), an albendazole-treated group, a native control group, and a pumpkin oil control group. Gas chromatography–mass spectrometry analysis of the pumpkin seed oil revealed a broad spectrum of biologically active compounds. The pumpkin seed oil treatment led to a significant reduction in the parasite burden, with a 75% decrease in adult worms and a 66% decrease in encysted larvae. Additionally, the infected animals treated with pumpkin oil exhibited a marked reduction in intestinal inflammation, characterized by a progressive increase in goblet cells. The number of encysted larvae in the diaphragm and muscle tissues was also significantly decreased. Furthermore, pumpkin seed oil treatment significantly reduced MMP-9 levels in both intestinal and muscular tissues, highlighting its potential to attenuate inflammation. Conclusion These findings underscore the effectiveness of pumpkin seed oil as anti-inflammatory and antiparasitic agent.