bimosiamose-disodium and Reperfusion-Injury

Preliminary studies in our laboratories indicate that a recently discovered synthetic drug, TBC-1269, acts as a multiple selectin blocker and provides protection against tissue damage in rats that are subjected to severe liver ischemia/reperfusion. Here, we report that this effect is dose and time dependent, with its effects acting through the modulation of tumor necrosis factor (TNF)-alpha and interleukin (IL)-10.. Mice subjected to 90 min of partial (70-80%) hepatic ischemia and 3 h of reperfusion were divided into eight groups (n=6/group): sham, ischemic control (IC), three groups of TBC-1269-treated animals at different concentrations (10, 20, 40, mg/kg) and another three groups of TBC-1269 given at 40 mg/kg at different times of administration: 15 min prereperfusion but after ischemia (no pretreatment), at the time of reperfusion, and at 15 min after reperfusion. The parameters measured at 3 h of reperfusion included liver function tests (alanine aminotransferase and aspartate aminotransferase), histopathology analysis and measurements using enzyme-linked immunosorbent assay in serum of TNF-alpha and IL-10. Statistical analysis included analysis of variance with P values of <0.05 for significance. Results were expressed as mean +/- SD.. The liver function tests showed statistically significant differences between the ischemic control group and both the sham group and the group treated with 40 mg/kg at the time of reperfusion (40@RP). These results correlated well with the histopathological analysis in that we found no difference in vacuolization, congestion, and necrosis between the 40@RP group and the sham group. The TNF-alpha and the IL-10 also reflected the protection observed in histopathology, with a decrease in TNF- alpha from the high levels observed in the IC (32 +/- 2.32 pg/mL) to a lower level of 8.5 +/- 4.04 mg observed in the 40@RP group, and an increment in the levels of the protective IL-10 from 2.8 +/- 2.9 pg/mL in the IC group versus 37.9 +/- 11.6 pg/mL in the 40@RP treated group (P<0.05). Lower doses and different times of administration of TBC-1269 did not show a protective effect. The IC group showed no difference in damage by histopathology or liver enzymes compared to the rest of the groups, except the 40@RP group.. In this work, we demonstrated that the small molecule multiple selectin inhibitor (TBC-1269) offered significant protection for the ischemic liver when given at 40 mg/kg at the time of perfusion. Lower doses and different times of administration did not show the optimal drug effect. The protection observed in the liver function tests (alanine aminotransferase and aspartate aminotransferase) and histopathology in this group was also reflected in the significant decrease in serum TNF-alpha and equally significant increase in serum protective IL-10.

Topics: Animals; Biphenyl Compounds; Disease Models, Animal; Dose-Response Relationship, Drug; Interleukin-10; Liver; Male; Mannose; Mannosides; Mice; Mice, Inbred C57BL; Reperfusion Injury; Selectins; Tumor Necrosis Factor-alpha

Experiments in rodents have demonstrated an important role for selectins in kidney ischemia-reperfusion injury (IRI). However, the relevance of this in larger mammals, as well as the impact on long-term structure and function is unknown. We tested the hypothesis that small molecule selectin ligand inhibition attenuates IRI, cellular inflammation, and long-term effects on renal interstitial fibrosis. We used a porcine model of kidney IRI and used Texas Biotechnology Corporation (TBC)-1269, a selectin ligand inhibitor. Renal function, tissue inflammation, and tubulointerstitial fibrosis development were evaluated up to 16 weeks. Both warm and cold ischemia models were studied for relevance to native and transplant kidney injury. Pigs treated with TBC-1269 during 45 min of warm ischemia (WI) showed significantly increased glomerular filtration rate compared to control animals. In pigs with severe IRI (WI for 60 min), TBC-1269 treatment during IRI significantly increased renal recovery. Cellular inflammation was strongly reduced, particularly influx of CD4 cells. Quantitative measurement of fibrosis by picrosirius red staining showed strong reduction in TBC-1269-treated groups. TBC-1269 also reduced cold IRI, inflammation, and fibrosis in kidneys preserved for 24 h at 4 degrees C and autotransplanted. The selectin ligand inhibitor TBC-1269 provides a novel and effective approach to attenuate IRI in both warm and cold ischemia in large mammals, in both short and long terms. Selectin ligand inhibition is an attractive strategy for evaluation in human kidney IRI.

Topics: Animals; Biphenyl Compounds; Disease Models, Animal; Fibrosis; Immunohistochemistry; Kidney; Ligands; Male; Mannose; Mannosides; Molecular Structure; Nephritis, Interstitial; Protective Agents; Reperfusion Injury; Selectins; Swine; T-Lymphocytes

Neonatal hearts have altered adhesion molecule interactions in response to ischemia-reperfusion. How this affects myocardial function is unknown.. Isolated, buffer perfused 0- to 2-day (newborn) and 2-week piglet hearts were first subjected to 20-minute global, normothermic ischemia, followed by 45 minutes of reperfusion during which 150 x 10(6) newborn or 2-week neutrophils were infused. In some hearts, an antibody to SLe(x) (CSLEX-1) was infused with neutrophils during reperfusion. Hemodynamic variables, including left ventricular developed pressure (LVDP), were recorded at timed intervals. Neutrophil CD-18, L-selectin, and SLe(x) contents were measured by flow cytometry.. Full recovery of LVDP was observed in newborn hearts receiving newborn or 2-week-old neutrophils. Recovery of LVDP was depressed (p < 0.01, ANOVA) in 2-week-old hearts receiving 2-week old, not newborn, neutrophils. Infusion of CSLEX-1 in 2-week-old hearts restored LVDP to baseline. Whereas flow cytometry showed higher (p < 0.01, Student's t test) CD-18 and L-selectin expression on newborn versus 2-week-old neutrophils, newborn neutrophils expressed lower (p < 0.01) SLe(x) levels.. Initial "loose" neutrophil-endothelial selectin interactions are a necessary prelude to "firm" adhesion and reperfusion injury. Operations performed soon after birth may be better tolerated than when surgery is delayed; anti-SLe(x) preparations may prove beneficial when performing cardiac procedures on older infants.

Topics: Animals; Animals, Newborn; Antibodies, Monoclonal; Biphenyl Compounds; CD18 Antigens; Chemotaxis, Leukocyte; Coronary Circulation; Heart Rate; Hemodynamics; L-Selectin; Lewis X Antigen; Mannose; Mannosides; Neutrophils; Reperfusion Injury; Swine

The selectin family of adhesion molecules plays a key role in the neutrophil-mediated injury observed after ischemia and reperfusion. In our study, we investigated the effects of TBC-1269, a novel small-molecule, nonoligosaccharide inhibitor of P-, E-, and L-selectin binding, in the liver inflammatory response after 90 minutes of warm ischemia.. Total liver ischemia was produced in Sprague-Dawley rats for 90 minutes using an extracorporeal portosystemic shunt. The animals were divided into five groups including: the sham (group 1), ischemic control (group 2) receiving only the vehicle, and the treated groups receiving TBC-1269 at a dose of 25 mg/kg at different times of administration: 15 minutes before reperfusion (group 3), at reperfusion (group 4), and 15 minutes after reperfusion (group 5). The following indices were analyzed: 7-day survival, liver injury tests, liver tissue myeloperoxidase as an index of neutrophil infiltration, and liver histology.. TBC-1269 treated groups experienced a significant increase in survival compared with controls. Best overall survival, 70%, was observed when TBC-1269 (Texas Biotechnology Corporation, Houston, TX) was administered 15 minutes before reperfusion (p < 0.05). This group also showed a marked decrease (p < 0.05) in liver enzyme levels at 6 hours after reperfusion. Neutrophil migration was also significantly ameliorated (81%), as reflected by decreased myeloperoxidase levels. We observed improved histologic damage scores in the treated group compared with controls (p < 0.05).. A small-molecule selectin inhibitor (TBC-1269) had a protective effect in livers subjected to 90 minutes of warm hepatic ischemia and 6 hours of reperfusion by decreasing neutrophil infiltration, migration and subsequent tissue damage. The best protective effect was achieved when the compound was administered 15 minutes before reperfusion. These findings offer a new therapeutic alternative for protection against ischemia and reperfusion injury.

Topics: Animals; Antibodies, Monoclonal; Biphenyl Compounds; Disease Models, Animal; Inflammation; Liver; Mannose; Mannosides; Necrosis; Neutrophil Activation; Peroxidase; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Selectins