The Effect of Urinary Trypsin Inhibitor Against Neuropathic Pain in Rat Models

BACKGROUND: Nerve injury sometimes leads to chronic neuropathic pain associated with neuroinflammation in the nervous system. In the case of chronic neuropathic pain, the inflammatory and algesic mediators become predominant and result in pain hypersensitivity following nervous system damage. It is well known that urinary trypsin inhibitor (ulinastatin, UTI) has an anti-inflammatory activity. Recently, the neuroprotective action of UTI on the nervous system after ischemic injury has been reported. Thus, we evaluated the neuroprotective effect of ulinastatin in a rat model of neuropathic pain. METHODS: Neuropathic pain was induced with L5 spinal nerve ligation (SNL) in male Sprague-Dawley rats weighing 100-120 g. The rats were divided into 3 groups, with n = 8 in each group. The rats in the control group (group 1) were administered normal saline and those in group 2 were administered UTI (50,000 U/kg) intravenously through the tail vein for 3 days from the day of SNL. Rats in group 3 were administered UTI (50,000 U/kg) intravenously from the 5th day after SNL. The paw withdrawal threshold was measured using the von Frey test for 3 days starting from the 5th day after SNL. RESULTS: The paw withdrawal thresholds were significantly increased in the rats of group 2 compared to the other groups (P < 0.05). CONCLUSIONS: Ulinastatin, which was administered for 3 days after SNL, increased the paw withdrawal threshold and it could have a neuroprotective effect in the rat model of neuropathic pain.

Urinary trypsin inhibitor attenuates lipopolysaccharide-induced neutrophil activation / 대한마취과학회지

BACKGROUND: Urinary trypsin inhibitor (UTI), which is speculated to have anti-inflammatory effects, is one of serine protease inhibitors found in human urine and blood. The present study was conducted to clarify the effect of urinary trypsin inhibitor (UTI) on human neutrophil activation and its intracellular signaling mechanism in vitro. METHODS: To assess the possible interactions between UTI and lipopolysaccharide (LPS) in neutrophil activation, neutrophils from human blood were incubated with varying concentrations of UTI (1, 10, 100, 1,000 and 10,000 U/ml) plus LPS (100 ng/ml) or LPS alone in 24-well plates (5 x 106 cells/well). We measured protein levels for interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-alpha) using enzyme-linked immunosorbent assay (ELISA) kits after 4 hours of incubation period. To elucidate the intracellular signaling pathway, we also measured the levels of phosphorylation of p38, ERK1/2 and JNK via Western blot analysis. Moreover, the nuclear levels of nuclear factor-kappa B (NF-kappaB) were determined with electrophoretic mobility shift assays (EMSA). RESULTS: UTI decreased the expression of inflammatory cytokines, including TNF-alpha and IL-6, and activation of intracellular signaling pathways, such as JNK, but not P38, ERK1/2 and nuclear translocation of NF-kappaB. CONCLUSIONS: UTI can attenuate LPS-induced neutrophil responses and may partially contribute to the treatment of neutrophil-mediated inflammatory diseases.

Effect of ulinastatin on cytokine reaction during gastrectomy / 대한마취과학회지

BACKGROUND: Inflammation plays an important role in the postoperative morbidity of organs, which is related to the activation of pro-inflammatory and anti-inflammatory cytokines. Ulinastatin (Urinary trypsin inhibitor, UTI) is a serine protease inhibitor found in human urine or serum that inhibits the activation of human leukocyte elastase. This study examined the effect of UTI on the inflammation response in patients undergoing a gastrectomy. METHODS: Thirty patients scheduled to undergo a gastrectomy were divided into two groups as follows: Control group (untreated, n = 15) and UTI group (100,000 units of UTI were continuously injected intravenously for 2 hours, n = 15). Arterial blood was sampled before surgery (T0), 10 minutes after its onset (T1), at its end (T2), and 1 hour after surgery (T3) to measure the level of cytokines. RESULTS: Both the control and treatment groups had higher interleukin (IL)-6 levels at T2 and T3 than T0, and the level increased with time. However, the increase was smaller in the treatment group. The IL-8 levels were not activated significantly in any of the groups. CONCLUSIONS: UTI inhibits the secretion of IL-6, which is an inflammatory cytokine produced after a gastrectomy. This shows that UTI can decrease the inflammation reaction caused by surgical stress.

The Effects of Urinary Trypsin Inhibitor on the Outcomes of Severe Sepsis and Septic Shock Patients

PURPOSE: We purposed to determine the effects of urinary typsin inhibitor (ulinastatin) on the outcomes of severe sepsis and septic shock patients. METHODS: This is a prospective case control study of severe sepsis and septic shock patients who visited emergency department of university hospital from January 2005 to June 2008. For study group, 100,000 U of ulinastatin was initially infused and then additional infusions of ulinastatin were determined by the mean arterial pressure. We compared the predicted mortality and the actual in-hospital mortality between the ulinastatin group and the control group. We also compared the improvement of the SOFA score according to time between the groups. RESULTS: There were 43 patients in the ulinastatin group and 126 patients in the control group. The predicted mortality and the actual mortality of the ulinastatin group were 31.2% and 18.6%, respectively. The predicted and actual mortalities of the control group were 33.1% and 27.0%, respectively. The improvement of the SOFA score for the ulinastatin group was 6.8+/-3.9 and 5.0+/-4.5 at 0 and 24 hours (p<0.001), 6.5+/-3.7 and 3.9+/-4.3 at 0 and 48 hours (p<0.001) and, 6.3+/-3.6 and 3.0+/-4.1 at 0 and 72 hours (p<0.001). For the control group, the change of the SOFA score was 4.9+/-2.9 and 5.8+/-4.1 at 0 and 24 hours (p=0.003), 5.0+/-2.8 and 5.1+/-4.2 at 0 and 48 hours (p=0.760) and, 4.8+/-2.7 and 4.34.1 at 0 and 72 hours (p=0.105). CONCLUSION: The ulinastatin group showed significantly lower mortality than the predicted mortality and the ulinastatin group's SOFA score was improved in the early hospital days.

Inhibitory effect of urinary trypsin inhibitor on IL-6 expression in severe acute pancreatitis in rats / 第三军医大学学报

Objective To explore the role of human urinary trypsin inhibitor(UTI) in severe acute pancreatitis (SAP). Methods UTI was highly purified from the urine of patients with acute pancreatitis by column chromatography. SAP models were established by injection of 5% sodium taurocholate(NaTc) at dose of 0.1 ml/100 g weight under pancreatic capsule. The interleukin 6(IL 6) level in each group was measured by ELISA. Results ① The highly purified UTI with high yield was harvested. ② SAP models were established successfully. ③ The level of IL 6 in SAP group was significantly higher than that in NC, but in UTI group, the increase was not obvious. Conclusion The alleviating effect of UTI on the pathological damage degree in rats with SAP is related to the inhibitive effect of UTI on the expression of IL 6.

Inhibitory effect of urinary trypsin inhibitor on protease activity in the colonic tissues of ulcerative colitis rats / 重庆医科大学学报

Objective:To determine the effect of urinary trypsin inhibitor(UTI)on protease activity in the colonic tissues of ulcerative colitis(UC) rats.Methods:The experimental UC was induced in Wistar rats by enema with 0.25ml of 18% 2,4-dinitrobenzenesulfonic acid (DNBS) dissolved in ethanol.UTI was injected into the rats through the caudal vein.Protease activity present in colonic tissues was determined on gelatin zymograms.Results:The protease activity which presented in colonic tissues of the group treated with UTI was only 1/3 that of the control.Conclusion:The protease activity decreased significantly after we treated the rats with UTI.