interleukin-8 and sivelestat

Patients undergoing lobectomy are at risk of developing acute lung injury resulting from one-lung ventilation (OLV) during surgery. We investigated the morphological and functional behavior of neutrophils in patients who underwent lobectomy and assessed the ability of sivelestat to inhibit neutrophil activity. This was a blinded randomized study. Sixteen patients who underwent lobectomy were given intravenous sivelestat (n = 8) or intravenous saline (n = 8). We studied the cytoskeletal rearrangements of circulating neutrophils by determining the localization of filamentous actin (F-actin). Pulmonary oxygenation was evaluated by measuring the partial pressure of arterial oxygen. We found that the number of circulating, F-actin-rimmed neutrophils increased during OLV and after lung re-expansion. Our results suggest that, in addition to the surgical procedure and OLV, re-expansion of the remaining lung after lobectomy increases the neutrophil activation levels. Furthermore, administration of sivelestat limited neutrophil activation and improved pulmonary oxygenation in our patients.

Topics: Actin Cytoskeleton; Actins; Acute Lung Injury; Aged; Aged, 80 and over; Female; Glycine; Humans; Interleukin-8; Lung; Male; Malondialdehyde; Middle Aged; Neutrophils; One-Lung Ventilation; Oxygen; Serine Proteinase Inhibitors; Sulfonamides; Thoracic Surgical Procedures

Post-operative pulmonary complications such as systemic inflammatory response syndrome (SIRS), acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are strongly associated with morbidity and mortality after esophagectomy. Post-operative administration of sivelestat sodium hydrate (sivelestat), a selective inhibitor of neutrophil elastase (NE), has been shown to improve the post-operative clinical course after esophagectomy. This study aimed to evaluate the effect of prophylactic administration of sivelestat on bronchial inflammatory responses. We randomized 24 patients into two groups. One group received 0.2 mg/kg/h sivelestat from the induction of anesthesia to post-operative day 1 (sivelestat group) and the other group received the same amount of physiological saline (control group). Bronchial alveolar epithelial lining fluid (ELF) samples were obtained from both groups at the induction of anesthesia and at the end of surgery. The serum and ELF levels of interleukin (IL)-6 and IL-8 were measured by enzyme-linked immunosorbent assay, and NE activity was spectrophotometrically determined using the same samples. Although IL-6 levels in the ELF significantly increased at the end of surgery compared with the pre-operative levels in both groups, the IL-8 levels and NE activity did not significantly increase at the end of the surgery compared to the corresponding pre-operative values in the sivelestat group. Moreover, IL-8 levels and NE activity in the ELF were significantly reduced at the end of surgery in the sivelestat group compared with corresponding values in the control group. The durations of ALI and ARDS were apparently shorter in the sivelestat group and the duration of SIRS was significantly shorter in the sivelestat group compared to the control group. We demonstrated that prophylactic use of sivelestat mitigated bronchial inflammation by suppressing NE activity and IL-8 levels in the ELF and shortened the duration of SIRS after transthoracic esophagectomy.

Topics: Acute Lung Injury; Aged; Bronchitis; Esophagectomy; Female; Glycine; Humans; Interleukin-6; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Postoperative Complications; Premedication; Respiratory Distress Syndrome; Respiratory Function Tests; Serine Proteinase Inhibitors; Sulfonamides; Systemic Inflammatory Response Syndrome; Treatment Outcome

Cardiopulmonary bypass (CPB) has been implicated as a cause of acute lung injury (ALI) in cardiac surgical patients. We used a bronchoscopic microsampling (BMS) probe to examine alveolar biochemical constituents and evaluated the effect of sivelestat sodium hydrate, a novel synthesized polymorphonuclear (PMN) neutrophil elastase inhibitor, on ALI induced by CPB. Twelve patients undergoing aortic valve replacement were treated with either sivelestat 0.2 mg/kg/h (sivelestat group, n=6) or 0.9% saline (control group, n=6) from the start of surgery. Samples were collected by the BMS probe at three time points: after tracheal intubation, 1 h after CPB introduction, and 3 h after CPB termination. Pulmonary function was assessed perioperatively. There were no differences in baseline characteristics. The concentration of PMN elastase was significantly suppressed in the sivelestat group, compared with the control group (P=0.001). The sivelestat group also had lower levels of interleukin-6 and interleukin-8. Alveolar-arterial oxygen difference markedly increased, and a worsening of the PaO(2)/FiO(2) ratio indicated severe impairment after CPB. However, sivelestat attenuated the pattern of physiological deterioration of gas exchange. Sivelestat may attenuate neutrophil elastase or proinflammatory cytokines, and improve pulmonary dysfunction in patients undergoing CPB.

Topics: Acute Lung Injury; Aged; Anti-Inflammatory Agents; Aortic Valve; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Bronchoscopy; Cardiopulmonary Bypass; Female; Glycine; Heart Valve Prosthesis Implantation; Humans; Inflammation Mediators; Infusions, Intravenous; Interleukin-6; Interleukin-8; Leukocyte Elastase; Lung; Male; Middle Aged; Proteinase Inhibitory Proteins, Secretory; Pulmonary Gas Exchange; Sulfonamides; Time Factors; Treatment Outcome

We evaluated the effect of sivelestat sodium (SiV), a novel synthesized polymorphonuclear (PMN) elastase inhibitor, on acute lung injury (ALI) caused by cardiopulmonary bypass (CPB).. Fourteen patients who underwent cardiopulmonary surgery using CPB, followed by the development of both systemic inflammatory response syndrome (SIRS) and ALI, were treated with either 0.2 mg/kg per hour SiV (SiV group, n = 7) or saline (control group, n = 7) for 4 days from the time of arrival in the intensive care unit.. The SiV group had a significantly lower ratio of serum PMN elastase and interleukin (IL)-8, a significantly lower ratio of the respiratory index, and a significantly higher ratio of PaO(2)/FiO(2) after 24 h of treatment than the control group.. Sivelestat sodium suppressed the production of PMN elastase and IL-8, resulting in improved respiratory function in patients with ALI caused by CPB.

Topics: Acute Disease; Aged; Cardiopulmonary Bypass; Female; Glycine; Humans; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Perfusion; Respiratory Distress Syndrome; Serine Proteinase Inhibitors; Sulfonamides; Systemic Inflammatory Response Syndrome

Sivelestat sodium hydrate (sivelestat) is a selective inhibitor of polymorphonuclear leukocyte elastase (PMN-E). We administered sivelestat to patients with septic acute lung injury (ALI) to examine its usefulness. The primary endpoints in the study were the duration of artificial ventilation and pulmonary oxygenation ability, and the secondary endpoints were mortality and the concentrations of PMN-E, SP-D, TNF-alpha and IL-8 in blood. In the sivelestat group, the duration of artificial ventilation, pulmonary oxygenation ability, and the blood PMN-E, SP-D, TNF-alpha and IL-8 concentrations decreased significantly. Administration of sivelestat was found to reduce alveolar dysfunction and improve respiratory function, and it was suggested that early administration might be useful.

Topics: Acute Disease; Adolescent; Adult; Aged; Glycine; Humans; Interleukin-8; Leukocyte Elastase; Leukocytes; Middle Aged; Oxygen; Pulmonary Circulation; Pulmonary Surfactant-Associated Protein D; Respiration, Artificial; Serine Proteinase Inhibitors; Sulfonamides; Survival Analysis; Systemic Inflammatory Response Syndrome; Treatment Outcome; Tumor Necrosis Factor-alpha

The purpose of this study is to examine the effects of neutrophil elastase inhibitor on postoperative cytokine levels in patients after esophagectomy.. Fifteen patients were divided into two groups; group 1: 9 patients receiving neutrophil elastase inhibitor (0.2 mg x kg(-1) x hr(-1) from the admission to ICU to extubation), group 2: 6 patients as a control. We measured neutrophil elastase activity, interleukin 1-beta, interleukin 6 and interleukin 8 preoperatively, just after the admission to ICU, and 24, 48 and 72 hours after the surgery.. There were significant differences in neutrophil elastase activity and interleukin 8 in group 1 24 hours after the surgery, compared with those in group 2. The time necessary for mechanical ventilation in group 1 was shorter than that in group 2 (group 1: 44.7 +/- 24.7 hrs, group 2: 112.8 +/- 90.3 hrs, P = 0.048).. The administration of neutrophil elastase inhibitor may be useful for patients after esophagectomy to reduce overexpression of plasma cytokine levels after surgery.

Topics: Cytokines; Esophageal Neoplasms; Esophagectomy; Glycine; Humans; Interleukin-8; Leukocyte Elastase; Lung Diseases; Postoperative Complications; Postoperative Period; Respiration, Artificial; Serpins; Sulfonamides; Time Factors

Patients with severe burns often develop acute lung injury (ALI), systemic inflammatory response syndrome (SIRS) often complicates with ALI. Sivelestat sodium hydrate is an effective drug against ALI. However, the mechanisms of this beneficial effect are still poorly understood. In the current study, we evaluate the effects of sivelestat sodium hydrate on systemic and local inflammatory parameters (neutrophil elastase [NE], interleukin [IL]-8, matrix metalloproteinase [MMP] 2 and 9) in a rat model of severe burns and ALI. And to analyze the correlations between expression of NE and IL-8 and acute lung injury.. 48 Sprague-Dawley (SD) rats were divided into 3 groups: normal control group, severe burns injury group and severe burns treated with sivelestat sodium hydrate group (SSI). The lung water content and PaO2 were detected in each group. Pathological manifestations in each group were observed for pathology scoring in SD rats with acute lung injury. ELISA was used for detecting expression of NE and IL-8 in serum and BAL specimens of SD rats in each group. RT-PCR was used to detect mRNA expression of NE and IL-8 in lung tissues of each group. Western blotting was used for detecting protein expression of MMP-2 and MMP-9 in lung tissues of each group. SPSS 18.0 was used for statistical analysis.. The PaO2 was significantly increased after sivelestat sodium hydrate intravenous injection. Pathological score and water content of lung tissue were significantly decreased in SSI group compared with severe burns injury group, slightly higher than that normal control group. NE and IL-8 levels significantly decreased in serum, BAL and lung tissue specimens after sivelestat sodium hydrate intravenous injection; Expression of MMP-2 and MMP-9 were significantly up-regulated in severe burns group and showed no significantly changed after sivelestat sodium hydrate intravenous injection.. In a rat model of severe burns and ALI, administration of sivelestat sodium hydrate improved symptoms of ALI and significantly decreased inflammatory parameters NE and IL-8.

Topics: Acute Lung Injury; Animals; Blood Gas Analysis; Burns; Disease Models, Animal; Glycine; Inflammation Mediators; Interleukin-8; Leukocyte Elastase; Lung; Rats; Rats, Sprague-Dawley; Sulfonamides; Systemic Inflammatory Response Syndrome

To observe and study the effects of sivelestat on acute lung injury in dogs with severe burn-blast combined injury.. Thirty-two male beagle dogs of clean grade were divided into 4 groups: uninjured group (U), combined injury control group (CIC), combined injury+low dose of sivelestat group (CI+LS), combined injury+high dose of sivelestat group (CI+HS), with 8 dogs in each group. Except for the dogs in group U which were not injured, the dogs in the other 3 groups were inflicted with severe burn-blast combined injury. According to the Parkland formula, the dogs in groups U and CIC were infused with physiological saline, and the dogs in groups CI+LS and CI+HS received sivelestat with the dosage of 0.5 and 2.0 mg·kg(-1)·h(-1) respectively in addition. The 24 h continuous intravenous infusion was carried out for 2 days. At post injury hour (PIH) 6, CT scanning was conducted to observe the lung damage. At PIH 2, 6, 12, 24, and 48, mean arterial pressure (MAP), respiratory rate (RR), extra vascular lung water (EVLW), pulmonary vascular permeability index (PVPI), PaO2, and PaCO2 were measured; the contents of neutrophil elastase (NE), IL-8, and TNF-α were determined by ELISA. At PIH 48, all the dogs were sacrificed, and the lung tissues were harvested to measure the wet to dry lung weight ratio. The same examination was carried out in the dogs of the group U at the same time points. Data were processed with analysis of variance of repeated measurement and LSD test.. (1) CT images showed some exudative lesions in the dogs of groups CIC and CI+LS but not in the dogs of groups U and CI+HS. (2) No statistically significant differences were observed in MAP at each time point between every two groups (with P values above 0.05). The RR values in group U were significantly different from those of the other 3 groups at all time points (with P values below 0.05). The values of EVLW and PVPI in 3 combined injury groups were significantly different from those in group U at PIH 6, 12, 24, and 48 (with P values below 0.05). The values of RR and EVLW in group CI+LS were significantly different from those in group CI+HS at PIH 12, 24, and 48 (with P values below 0.05). The values of PVPI in group CI+LS were significantly different from those in group CI+HS at PIH 24 and 48 (with P values below 0.05). (3) The levels of PaO2 and PaCO2 showed significant differences between group U and the other 3 groups at each time point (with P values below 0.05). The levels of PaO2 in group CI+LS were significantly different from those in CI+HS group at PIH 12, 24, and 48 (with P values below 0.05). The level of PaCO2 showed significant differences between group CI+LS and group CI+HS at PIH 24 and 48 (with P values below 0.05). (4) The contents of NE (except for PIH 2), TNF-α, and IL-8 showed significant differences between group U and the other 3 groups at each time point (P < 0.05 or P < 0.01). At PIH 2, 6, 12, 24, and 48, the contents of NE in groups U, CIC, CI+LS, and CI+HS were respectively (69 ± 21), (83 ± 24), (80 ± 20), (75 ± 17), (72 ± 27) pg/mL; (66 ± 24), (196 ± 20), (231 ± 26), (252 ± 25), (266 ± 22) pg/mL ; (71 ± 22), (180 ± 27), (214 ± 21), (194 ± 24), (218 ± 20) pg/mL; (68 ± 22), (136 ± 24), (153 ± 22), (146 ± 26), (150 ± 28) pg/mL. NE values in group CI+HS were statistically different from those in groups CIC and CI+LS at PIH 6, 12, 24, and 48 (with P values below 0.05). The contents of TNF-α in group CI+LS were statistically different from those in groups CIC and CI+HS at PIH 24 and 48 (with P values below 0.05). The contents of IL-8 in group CI+LS were statistically different from those in group CI+HS at PIH 24 and 48 (with P values below 0.05). (5) At PIH 48, the wet to dry lung weight ratio of group CIC was statistically different from that in group CI+LS or group CI+HS (with P values below 0.05); there was also difference between group CI+LS and group CI+HS (P < 0.05).. Sivelestat, especially in a high dose, exerts a protective effect in acute lung injury after burn-blast combined injury through improving the index of blood gas analysis, ameliorating pulmonary edema, and lowering the production of pro-inflammatory mediators.

Topics: Acute Lung Injury; Animals; Blood Gas Analysis; Burns; Capillary Permeability; Dogs; Extravascular Lung Water; Glycine; Infusions, Intravenous; Interleukin-8; Male; Pulmonary Edema; Serine Proteinase Inhibitors; Sulfonamides; Tumor Necrosis Factor-alpha

Neutrophil elastase (NE) takes part in the pathogenesis of acute lung injury. However, its role in lung injury of burn-blast combined injury is unclear. Our objective was to assess the role of NE, and effect of sivelestat, a specific NE inhibitor, in lung injury induced by burn-blast combined injury in rats.. One hundred and sixty male Sprague-Dawley rats were randomly subjected to burn-blast combined injury (BB) group, burn-blast combined injury plus sivelestat treatment (S) group or control (C) group. Blood gas, protein concentration and NE activity in bronchoalveolar lavage fluid (BALF), pulmonary myeloperoxidase (MPO) activity, serum concentrations of TNF-α and IL-8, etc. were investigated from 0 h to 7 d post-injury.. In BB group, PaO2 decreased, while NE activity in BALF, total protein concentration in BALF, pulmonary MPO activity and W/D ratio, serum concentrations of TNF-α and IL-8 increased with neutrophil infiltration, progressive bleeding and pulmonary oedema. Compared with BB group, sivelestat treatment decreased the NE activity and ameliorated the above indexes.. Sivelestat, exerts a protective effect in lung injury after burn-blast combined injury through inhibiting NE activity to decrease pulmonary vascular permeability, neutrophil sequestration, and production of TNF-α and IL-8.

Topics: Animals; Blast Injuries; Bronchoalveolar Lavage Fluid; Burns; Carbon Dioxide; Disease Models, Animal; Glycine; Interleukin-8; Leukocyte Elastase; Lung Injury; Male; Oxygen; Partial Pressure; Proteinase Inhibitory Proteins, Secretory; Rats; Rats, Sprague-Dawley; Serine Proteinase Inhibitors; Sulfonamides; Tumor Necrosis Factor-alpha

Intrauterine infection and inflammation are recognized as major contributors to the onset of preterm labor. We describe two cases of severe preterm labor with bulging membrane that were treated by intravenous injection of Sivelestat, a neutrophil elastase inhibitor. Ritodrine hydrochloride and magnesium sulfate were intravenously administered for tocolysis, and ampicillin was provided as an antibiotic. Urinary trypsin inhibitor (UTI) was administered transvaginally. Sivelestat was infused intravenously at 4.8 mg kg(-1) day(-1) through the maternal vein. No side effects were observed. Levels of interleukin (IL)-6 and IL-8 in amniotic fluid decreased, and gestations were prolonged without complications for >1 week. Two healthy infants were delivered. Our experience suggests that multidrug therapy with Sivelestat offers a new therapeutic strategy for preterm labor, but further investigations of the indications, administration period and dosage are required.

Topics: Adult; Amniotic Fluid; Female; Fetal Membranes, Premature Rupture; Glycine; Humans; Injections, Intravenous; Interleukin-6; Interleukin-8; Magnesium Sulfate; Obstetric Labor, Premature; Pregnancy; Ritodrine; Serine Proteinase Inhibitors; Sulfonamides; Tocolytic Agents; Young Adult

Sivelestat sodium hydrate is a selective inhibitor of neutrophil elastase, which is effective in acute lung injury associated with systemic inflammatory response syndrome. However, the effectiveness of sivelestat in sepsis has not been fully examined. In the present study, the effect of sivelestat on severe sepsis in a rat cecal ligation and puncture (CLP) model was investigated. Adult male Sprague-Dawley rats underwent CLP and were randomly divided into two groups: sivelestat-treated group and saline-treated controls. The serum concentrations of several inflammatory mediators were measured. Hematoxylin-eosin staining, and immunohistochemical staining for high-mobility group box chromosomal protein 1 (HMGB1), IL-8, and CD68 were performed on the lungs to assess pathological changes found 12 h after the CLP procedure. Treatment with sivelestat significantly improved the survival rate of the post-CLP septic animals (P = 0.030). Sivelestat also induced a significant reduction in serum IL-1beta (P = 0.038) and IL-10 (P = 0.008) levels in these CLP rats. Serum HMGB1 levels had no significant difference between the sivelestat-treated and the control group. The lungs from sivelestat-treated rats exhibited less severe pathological changes and decreased the numbers of HMGB1, IL-8, and CD68-positive cells (P < 0.001). Sivelestat significantly improved survival rate of rats with clinically relevant sepsis, possibly by attenuating sepsis-induced systemic inflammatory response and lung injury. This may explain the implicated health benefits of sivelestat in reducing morbidity and mortality from sepsis.

Topics: Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cecum; Disease Models, Animal; Glycine; HMGB1 Protein; Inflammation; Interleukin-8; Ligation; Lung; Male; Proteinase Inhibitory Proteins, Secretory; Rats; Rats, Sprague-Dawley; Sepsis; Sulfonamides; Survival Analysis

The goal of the study was to examine the effects of sivelestat sodium hydrate (sivelestat), a neutrophil elastase inhibitor, on production of cytokines in granulocytes and monocytes, using flow cytometry after cytokine staining in whole blood culture.. Blood samples were collected from healthy volunteers. Vehicle (control group), lipopolysaccharide (LPS) (LPS group), or LPS + sivelestat (sivelestat group) were added to the whole blood, followed by addition of a protein transport inhibitor in each group. After incubation, staining for cytokines retained in the cells was performed by addition of an anti-interleukin 8 (IL-8) or anti-tumor necrosis factor-α (TNF-α) antibody. The cells were then analyzed using flow cytometry.. Granulocytic production of IL-8 induced by 1 ng/ml LPS was significantly (P < 0.05) inhibited by treatment with 1 μg/ml sivelestat, and upregulation of IL-8 by 10 ng/ml LPS was also significantly (P < 0.05) suppressed by 1 and 10 μg/ml sivelestat. Addition of 10 or 100 μg/ml sivelestat significantly (P < 0.05) inhibited the production of TNF-α from granulocytes induced by 10 ng/ml LPS. Sivelestat did not significantly inhibit LPS-induced monocytic production of TNF-α and IL-8.. Suppression of granulocytic production of IL-8 and TNF-α by sivelestat suggests that this drug may be useful for treatment of morbid conditions involving IL-8 and TNF-α at onset.

Topics: Acute Lung Injury; Flow Cytometry; Glycine; Granulocytes; Humans; In Vitro Techniques; Interleukin-8; Lipopolysaccharides; Monocytes; Proteinase Inhibitory Proteins, Secretory; Sulfonamides; Tumor Necrosis Factor-alpha

Neutrophil elastase probably contributes to the development of acute lung injury after cardiopulmonary bypass (CPB) in patients with infection or shock. We evaluated whether pretreatment with sivelestat sodium hydrate, a neutrophil elastase inhibitor (EI), can prevent acute lung injury caused by CPB.. Rabbits were assigned four groups: CPB for 60 minutes, control CPB group; low-dose lipopolysaccharide (LPS) administration without CPB, LPS group; CPB after lipopolysaccharide administration, LPS+CPB group; or preparation with continuous infusion of sivelestat and CPBs after lipopolysaccharide administration, EI group. Blood samples to determine blood gas concentration, plasma elastase activity, and plasma interleukin-8 levels were obtained. Histopathologic examinations of the lung were performed.. The decreased arterial oxygen pressure at the end of CPB was observed in the LPS+CPB group only, but was suppressed in the EI group (p < 0.01). Elastase activity was markedly elevated at 120 minutes after CPB, and interleukin-8 levels were markedly elevated at 180 minutes in the LPS+CPB group but were much lower (p < 0.05) in the EI group. Histopathology demonstrated accumulation of polymorphonuclear neutrophils in bronchoalveolar areas in the LPS+CPB group (p < 0.01). Pulmonary myeloperoxidase activity was significantly lower in the LPS+CPB group than in the other groups (p < 0.01). These changes were minimal in the EI group.. The combination of low dose LPS+60 minutes of CPB, but neither intervention alone, produced evidence of acute lung injury in a rabbit model. This did not occur when the animals were pretreated with sivelestat.

Topics: Animals; Cardiopulmonary Bypass; Glycine; Interleukin-8; Leukocyte Elastase; Lipopolysaccharides; Neutrophils; Peroxidase; Rabbits; Respiratory Distress Syndrome; Sulfonamides

Although liberalization of donor criteria may be one of the solutions to the current serious lung donor shortage, the use of non-standard donor lungs would increase the risk of post-operative complications. In the present study, we investigated the effect of sivelestat, a neutrophil elastase inhibitor, on reperfusion injury of a donor lung that was harvested from endotoxin-primed animals in a rat lung transplantation model.. Donor rats received an intraperitoneal injection of Escherichia coli endotoxin (5 mg/kg) 2 hours before lung harvesting. The donor lungs were flushed with an organ preservation solution with or without sivelestat (300 microg/ml), and the left lung was immediately transplanted to the recipient by the cuff technique.. Endotoxin priming did not cause significant lung injury before harvesting. Although these lungs looked normal macroscopically, they were found to contain numerous neutrophils in the alveolar capillaries, even after lung flushing. There was no significant difference in the neutrophil count between the lungs flushed with and without sivelestat. The endotoxin-primed donor lung without sivelestat treatment became edematous immediately after reperfusion. In addition, the recipient's native right lungs were also pathologic. Treatment with sivelestat significantly reduced injury in both the donor and the recipient's native lungs. Treatment with sivelestat also inhibited the increase in tumor necrosis factor-alpha and cytokine-induced neutrophil chemoattractant-1 levels in the recipient circulation after reperfusion.. We conclude that sivelestat could reduce lung injury after transplantation by inhibiting the deleterious burst of inflammatory reactions that are initiated by reperfusion of the lungs from endotoxin-primed rats.

Topics: Animals; Cytokines; Endotoxins; Escherichia coli; Glycine; Injections, Intraperitoneal; Interleukin-8; Leukocyte Elastase; Lung; Lung Transplantation; Male; Neutrophils; Pneumonia; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Sulfonamides; Tissue and Organ Harvesting; Tumor Necrosis Factor-alpha

The alveolar epithelial cell type II (AEC-II) is itself able to amplify lung inflammation by producing inflammatory cytokines and chemokines, leading to the activation and recruitment of phagocytes. Sivelestat, a new neutrophil elastase inhibitor, has been shown to attenuate acute lung injury in animal experiments. In the current study, we assessed the effects of sivelestat on the production of chemokines from cultured A549 cells, a human AEC-II-like cell line.. A549 cells were stimulated with endotoxin or tumor necrosis factor-alpha in the presence of sivelestat (1-100 microg x ml(-1)). Culture supernatant levels of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) were determined by enzyme-linked immunosorbent assay. The expression of IL-8 and MCP-1 mRNAs in stimulated A549 cells in the presence of sivelestat (100 microg x ml(-1)) was quantified by real-time polymerase chain reaction.. Sivelestat, at 100 microg x ml(-1) reduced the accumulation of IL-8 and MCP-1 in the culture medium. The high dose of sivelestat significantly inhibited the expression of IL-8 mRNA in A549 cells. The drug also decreased MCP-1 mRNA expression, although not significantly.. These data suggest that a high dose of sivelestat regulates the production of IL-8 and MCP-1 in AEC-II.

Topics: Cells, Cultured; Chemokine CCL2; Dose-Response Relationship, Drug; Endotoxins; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Glycine; Humans; In Vitro Techniques; Interleukin-8; Polymerase Chain Reaction; Pulmonary Alveoli; Respiratory Mucosa; RNA, Messenger; Serine Proteinase Inhibitors; Sulfonamides; Time Factors; Tumor Necrosis Factor-alpha

Neutrophil is a major focus in efforts to ameliorate the systemic inflammatory response associated with cardiopulmonary bypass. Neutrophil elastase is a powerful proteolytic enzyme, and plays a pivotal role in the development of the inflammatory response. This study assesses the inhibitory effects of sivelestat, a highly specific neutrophil elastase inhibitor, on elastase levels, cytokine production, and the functional changes of neutrophils in a simulated extracorporeal circulation model.. Simulated recirculation was established by recirculating heparinized (3.75 U/mL) human blood for 120 minutes in an oxygenator and a roller pump circuit with and without 100 micromol/L of sivelestat (n = 7 for each group). Neutrophil elastase and interleukin-8 were measured with an enzyme immunoassay. Neutrophil deformability was evaluated by simulated microcapillaries. The neutrophil F-actin and the expression of CD11b and L-selectin were measured by flow cytometry.. Sivelestat reduced both neutrophil elastase levels (p = 0.0006) and interleukin-8 production (p < 0.0001) at 120 minutes of recirculation. Sivelestat also significantly preserved neutrophil deformability (p = 0.017) and reduced F-actin expression (p = 0.0037). The drug did not modulate the changes of CD11b or L-selectin.. This study suggests that specific elastase inhibition with sivelestat could be a feasible therapeutic strategy for patients undergoing cardiopulmonary bypass to attenuate neutrophil-derived inflammatory response and organ injuries.

Topics: Actins; CD11b Antigen; Cell Adhesion Molecules; Chemotactic Factors; Chemotaxis, Leukocyte; Extracorporeal Circulation; Glycine; Humans; Inflammation Mediators; Interleukin-8; L-Selectin; Leukocyte Elastase; Models, Cardiovascular; Neutrophils; Serine Proteinase Inhibitors; Sulfonamides

In view of a cytoprotective effect of elastase inhibitor on chemokine-mediated tissue injury, we examined the neuroprotective effect of ONO-5046, a specific inhibitor of neutrophil elastase, in rats with spinal cord injury. Standardized spinal cord compression markedly increased cytokine-induced neutrophil chemo-attractant (CINC)-1 mRNA and protein. Their increases correlated with neurologic severity of injured rats. Immunohistochemically, CINC-1 protein was detected sequentially in vascular endothelial cells at 4 h, in perivascular neutrophils at 8 h, and in neutrophils infiltrating into cord substance at 12 h. Pretreatment with ONO-5046 (50 mg/kg) markedly ameliorated motor disturbance in injured rats, and reduced CINC-1 protein and mRNA expression. ONO-5046 also significantly reduced the increase of neutrophil accumulation or infiltration estimated by myeloperoxidase activity, and the extent of vascular permeability by Evans blue extravasation in the injured cord segment in comparison to control animals receiving vehicle. These results suggest that CINC-1 contributed to inflammation in rat spinal cord injury and ONO-5046 attenuated neurologic damage partly by blocking CINC-1 production of the chemoattractant, preventing neutrophil activation and vascular endothelial cell injury.

Topics: Animals; Blood-Brain Barrier; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Gene Expression; Glycine; Growth Substances; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Interleukin-8; Leukocyte Elastase; Motor Activity; Peroxidase; Rats; Rats, Wistar; Recovery of Function; RNA, Messenger; Serine Proteinase Inhibitors; Spinal Cord; Spinal Cord Injuries; Sulfonamides

Although neutrophil migration from the systemic circulation involves the beta2- (or CD18) integrin family, the existence of an alternative, CD18-independent route of neutrophil extravasation to tissues has been demonstrated in animal models. The molecular interactions involved in this alternative migratory route have not yet been characterized. The objective of this study was to assess the CD18-dependency of neutrophil migration across human endothelial cells from an organ known to support CD18-independent migration, the lung, with a view to establishing an in vitro model to facilitate study of CD18-independent migration. Neutrophil migration across human pulmonary artery endothelial cells (HPAECs) in response to three different chemoattractants, formylmethionyl leucylphenyl-alanine (FMLP), interleukin (IL)-8, and leukotriene (LT) B(4), was examined. Results demonstrated that a function-blocking antibody to CD18 decreased FMLP-stimulated migration by 71.7 +/- 4.4% (P < 0.001). In contrast, migration in response to LTB(4) was decreased by only 20.5 +/- 10.2% (P < 0.01), and no significant decrease was observed with migration to IL-8. Neutrophils that migrated to FMLP had 1.7-fold more surface CD11b/CD18 compared with nonmigrated neutrophils (P < 0.01), whereas this integrin complex was not significantly upregulated on neutrophils that had migrated to IL-8 or LTB(4). Further investigation of this migratory route indicated that it did not involve the beta1 integrins (CD29) or the endothelial selectins, E- or P-selectin, nor did it require the activity of either metalloproteinases or neutrophil elastase. These results indicate that neutrophil migration across HPAECs in vitro to IL-8 and LTB(4) is predominantly CD18-independent and provides a much-needed in vitro system for examination of the neutrophil-endothelial interactions involved in this alternative migratory route.

Topics: CD11 Antigens; CD18 Antigens; Cell Line; Cell Movement; Chemotactic Factors; Dose-Response Relationship, Drug; Endothelium, Vascular; Glycine; Humans; Hydroxamic Acids; Interleukin-8; L-Selectin; Leukotriene B4; Lung; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Permeability; Phenylalanine; Protease Inhibitors; Pyrazines; Sulfonamides; Thiophenes

Polymorphonuclear neutrophil elastase might contribute to postperfusion lung injury, so we evaluated the protective effect of ONO-5046*Na, a specific inhibitor of polymorphonuclear neutrophil elastase, against such an injury.. The study was done using 8 mongrel dogs that received ONO-5046*Na (15 mg/kg per hour) (group O) and 8 control dogs (group C), all of which had 1 hour of partial bypass and 5 hours of observation.. The respiratory index showed no significant changes in group O, but increased significant in group C (1.4+/-2.0 versus 5.1+/-4.7, p = 0.0047). Pulmonary extravascular water volume increased markedly in group C but only slightly in group O (group C 20.6+/-8.7, group O 11.2+/- 2.7 mL/kg; p = 0.0005). Blood concentrations of polymorphonuclear neutrophil elastase and interleukin-6 showed more than a tenfold increase in group C (PMN elastase, group C 12.9+/-12.8, group O 2.4+/-1.3 ng/mL; IL-b, group C 11.0+/-9.3, group O 2.9+/-3.8 pg/mL; p < 0.05) but were only slightly higher in group O. Histologic examination revealed interstitial and intraalveolar edema in group C, but group O was virtually normal.. ONO-5046*Na inhibits polymorphonuclear neutrophil elastase and maintains better pulmonary function, so it should reduce postperfusion lung injury.

Topics: Animals; Cardiopulmonary Bypass; Dogs; Extravascular Lung Water; Glycine; Hemodynamics; Interleukin-6; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Lung; Oxygen; Protective Agents; Pulmonary Circulation; Reperfusion Injury; Serine Proteinase Inhibitors; Sulfonamides; Vascular Resistance

The objective of our study was to elucidate the involvement of interleukin (IL)-8 in the hCG-induced rabbit ovulatory process. After administering hCG (100 IU i.v.), we examined myeloperoxidase (MPO) activity, which represents neutrophil accumulation; neutrophil elastase (NE) activity, which is an indicator of neutrophil activity; and levels of IL-8 in the ovaries. The maximal level of IL-8 was observed before MPO and NE activities reached a peak: production of IL-8, MPO, and NE activities peaked, respectively, at 4 h (5.58 +/- 0.88 pg/mg ovary, n = 13), 6 h (1.07 +/- 0.13 deltaA/min per gram ovary, n = 8), and 9 h (18.89 +/- 1.05 U/g ovary, n = 8). Anti-rabbit IL-8 antiserum given i.v. significantly reduced the maximal levels of hCG-induced MPO activity (antiserum vs. control; 0.34 +/- 0.04 vs. 1.12 +/- 0.11 deltaA/min per gram ovary, n = 14, p < 0.001) and NE activity (8.14 +/- 0.85 vs. 18.30 +/- 0.79 U/g ovary, n = 14, p < 0.001). The hCG-induced ovulation rate was significantly inhibited by the antiserum (50.5% vs. 83.9%, n = 14, p < 0.001). Intraperitoneal injection of 100 mg/kg of ONO-5046, a specific NE inhibitor, also attenuated the ovulation rate (ONO-5046 vs. vehicle; 56.0% vs. 74.0%, n = 14, p < 0.05). These findings clearly indicate that IL-8 has an important role in the hCG-induced ovulatory process through the accumulation and activation of neutrophils.

Topics: Animals; Chorionic Gonadotropin; Female; Glycine; Humans; Interleukin-8; Neutrophil Activation; Neutrophils; Ovary; Ovulation; Peroxidase; Rabbits; Serine Proteinase Inhibitors; Sulfonamides

Repeated intranasal administration of interleukin 8 (IL-8) induces bronchial hyperresponsiveness (BHR) accompanied by lower airway neutrophil accumulation (ANA) in guinea-pigs. Leukotriene B4 (LTB4) is a chemotactic factor for neutrophils. To elucidate whether LTB4 and neutrophil elastase are involved in the IL-8-induced BHR and ANA, the effects of a LTB4 antagonist (ONO-4057) and a neutrophil elastase inhibitor (ONO-5046) on the responses were examined. IL-8 (5 microg x kg[-1]) was administered intranasally to guinea-pigs twice weekly for 3 weeks. One day after the last administration, animals were anaesthetized and artificially ventilated through tracheal cannulae, and lateral pressure at the tracheal cannula (Pao) was measured as an overall index of airway responses to inhaled histamine. ONO-4057 (2 or 20 mg x kg[-1]) or ONO-5046 (30 or 300 mg x kg[-1]) was administered intraperitoneally 24 and 1 h before anaesthesia. ONO-4057, but not ONO-5046, significantly inhibited the IL8-induced BHR and ANA, assessed by bronchoalveolar lavage, in a dose-dependent manner. These findings suggest that interleukin 8 causes bronchial hyperresponsiveness and airway neutrophil accumulation in guinea-pigs in vivo. In part this appears to be due to release of leukotriene B4, whereas it may not be mediated by neutrophil elastase.

Topics: Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Movement; Glycine; Guinea Pigs; Immunosuppressive Agents; Interleukin-8; Leukocyte Elastase; Leukotriene B4; Male; Neutrophils; Phenylpropionates; Sulfonamides