Background: Intraperitoneal adhesions develop in up to 95% of patients following laparotomy, often accounting for significant long-term morbidity. Studies suggest that adhesions are reduced by mechanisms that upregulate fibrinolysis within the peritoneum. Statins promote fibrinolysis in the cardiovascular system and consequently may play a role in the prevention of adhesions. The aims of this study were to determine if statins reduce adhesion formation in vivo and to identify the mechanism of action in vitro. Methods: Adhesions were surgically induced in male Wistar rats (N=102) using a previously described ischemic button model. Rats received either vehicle (controls), lovastatin (30mg/kg), or atorvastatin (30mg/kg) as a single intraperitoneal dose at the time of laparotomy. Animals were sacrificed and adhesions were quantified at day 7. Peritoneal fluid and tissue were collected at day 1 to measure tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) by real-time PCR and ELISA. To assess the effects of statins on tissue healing, burst pressures were measured in anastomoses of the colon. The effects of lovastatin on tPA and PAI-1 production were measured in vitro in human mesothelial cells (HMC) in the presence or absence of mevalonate (MVA), geranylgeranyl-pyrophosphate (GGPP) and farnesyl-pyrophosphate (FPP), all intermediates in the cholesterol biosynthetic pathway downstream of HMG-CoA. The effect of a Rho protein inhibitor, exoenzyme C3 transferase, on tPA production was also determined. Results: The administration of both lovastatin and atorvastatin reduced adhesion formation by 26% and 58%, respectively (p<0.05) without affecting anastomotic burst pressure. tPA mRNA levels in peritoneal tissue and tPA activity in peritoneal fluid from lovastatin-treated animals were increased by 57% and 380%, respectively (p<0.05), while PAI-1 levels were unchanged. HMC incubated with either lovastatin or atorvastatin showed concentration-dependent increases in tPA production and decreases in PAI-1 production (p<0.05). These lovastatin-induced changes in tPA and PAI-1 production were significantly reversed by the addition of MVA, GGPP and FPP. The Rho protein inhibitor increased tPA production and rescued tPA production from the inhibitory effect of GGPP. Conclusion: These data suggest that statins administered within the peritoneum can upregulate local fibrinolysis, while the in vitro studies show that this effect may be mediated, in part, by intermediates of the cholesterol biosynthetic pathway that regulate Rho protein signaling. Therefore, statins may provide a viable strategy to prevent adhesion formation.