Introduction: Previous studies revealed anti-inflammatory and anti-spasmodic effects of Matricaria recutita. The objective of present study is to determine: 1) the effect of spray dried total extract of M. recutita collected in Golestan province of Iran on contractile phase of isolated jejunum of rabbit 2) the role of Cholinergic and Nitregic systems on anti-spasmodic effects of M. recutita.\ud Material &Methods: In first groups (N=6), the effects of increasing concentrations of plant extract on the contractile responses of jejunum of rabbits induced by cumulative addition of extracellular calcium and high K+ were studied. The role of the nitrergic and cholinergic systems was assessed in the presence of various concentrations of M. recutita (0.003–0.013mg/ml) and cholinerhic and nitrergic modulators at the second and third groups (N=12).\ud Results: Significant concentration-dependet inhibitory effects of M. recutita on contraction of smooth muscle caused by cumulative concentrations of calcium (0.03–3 mM) and to 50 mM Kcl was observed. Pretreatments with L-Name (100µm) and atropine (3µm) abrogated effectively the spasmolytic effect of M. recutita. Although, the M. recutita effect was significantly reduced in preparations pretreated with L-Argening (100mM), a precursor of Nitric Oxide production. Interestingly, contractile response of smooth muscle to Ach was completely reverted by M. recutita\ud Conclusion:Our results indicate that the anti-spasmodic effects of Matricaria recutita arise from its direct inhibitory effects on Ca2+ channels. however, modulation of cholinergic and nitregic pathway could be involved. These observations suggest that Matricaria recutita inhibit contractile elements of peristalsis by facilitating inhibitory role of nitrergic pathway