Anti-ischemic effect of selectin blocker through modulation of tumor necrosis factor-alpha and interleukin-10.
BACKGROUND
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
López-Neblina
Fernando
F
Surgery, Trauma and Molecular Biology, Borgess Research Institute, Kalamazoo, Michigan, USA.
Toledo-Pereyra
Luis H
LH
D000818:Animals; D001713:Biphenyl Compounds / Q000494:pharmacology*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D016753:Interleukin-10 / Q000378:metabolism*; D008099:Liver / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008358:Mannose / Q000031:analogs & derivatives; D008362:Mannosides / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000276:immunology / Q000473:pathology; D019087:Selectins / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D001713:Biphenyl Compounds; D008362:Mannosides; D019087:Selectins; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; C108700:bimosiamose disodium; D008358:Mannose