FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2021983732> ?p ?o ?g. }

• W2021983732 endingPage "1755" @default.
• W2021983732 startingPage "1749" @default.
• W2021983732 abstract "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°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. 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°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. Ischemic acute renal failure remains a major clinical problem with high morbidity and mortality in native kidneys, as well as with worse short- and long-term allograft function in allografts. A number of studies have examined the role of inflammatory cells in the pathogenesis of post-ischemic injury (reviewed by Friedewald and Rabb1.Friedewald J.J. Rabb H. Inflammatory cells in ischemic renal failure.Kidney Int. 2004; 66: 486-491Abstract Full Text Full Text PDF PubMed Scopus (347) Google Scholar). Leukocyte rolling on the endothelium, which precedes firm adhesion via CD11/CD18 and intercellular adhesion molecule (ICAM-1), is mediated by a class of adhesion molecules known as selectins, which consist of three different transmembrane receptors, E-, P-, and L-selectin.2.Lasky L. Selectins: interpreters of cell specific carbohydrates information during inflammation.Science. 1992; 259: 964-969Crossref Scopus (1151) Google Scholar E- and L-selectin are involved at the stage of leukocytes rolling and tethering to the vascular endothelium. L-selectin is constitutively expressed on leukocytes, whereas E-selectin is expressed on only activated endothelium. P-selectin is expressed on platelets and activated endothelial cells, and appears to have some inducers different from those of E-selectin.3.Chamoun F. Burne M. O'Donnell M. Rabb H. Pathophysiologic role of selectins and their ligands in ischemia reperfusion injury.Front Biosc. 2000; 5: 103-109Crossref PubMed Scopus (88) Google Scholar Interstitial inflammation is associated with the development of tubulointerstitial fibrosis, characterized by excess accumulation of extracellular matrix in the renal interstitium.4.Eitner F. Floege J. Novel insights into renal fibrosis.Curr Opin Nephrol Hypertens. 2003; 12: 227-252Crossref PubMed Scopus (33) Google Scholar Renal fibrosis is generally regarded as the dark side of tissue repair mechanisms and the ultimate injury that leads to chronic nephropathy. Selectin blockade has been demonstrated to attenuate ischemia-reperfusion injury (IRI) in different models.5.Seekamp A. Till G.O. Mulligan M.S. et al.Role of selectin in local and remote tissue injury following ischemia and reperfusion.Am J Pathol. 1994; 144: 592-598PubMed Google Scholar, 6.Springer T.A. Traffic signals for lymphocytes recirculation and leukocytes emigration: the multistep paradigm.Cell. 1994; 76: 301-314Abstract Full Text PDF PubMed Scopus (6395) Google Scholar, 7.Weyrich A.S. Ma X. Lefer D.J. et al.In vivo neutralization of P-selectin protects feline heart and endothelium in myocardial ischemia and reperfusion injury.J Clin Invest. 1993; 91: 2690-2692Crossref Scopus (448) Google Scholar, 8.Rabb H. Ramirez G. Saba S.R. et al.Renal ischemia–reperfusion injury in L-selectin deficient mice.Am J Physiol. 1996; 271: F408-F413PubMed Google Scholar However, blockade of one selectin can lead to compensatory inflammation conducted by the other selectins. A potent selectin antagonist, biosmanose Texas Biotechnology Corporation (TBC)-1269(1,6-Bis(3-(3-carboxymethylphenyl)-4-(2-α-D-mannnopyranosylloxy)-phenyl)hexane) is a small molecule antagonist of E-, P-, and L-selectin binding to their ligand, sialyl Lewis X. The structure of TBC-1269 reveals two mannose and two carboxylic acid groups in a configuration to mimic the dimeric configuration of sialyl-Lewis (Figure 1). TBC-1269 was demonstrated to mitigate post-ischemic renal damage in a rodent model during the first 72 h after renal ischemia.9.Nemoto T. Burne M.J. Daniels F. et al.Small molecule selectin ligand inhibition improves outcome in ischemic acute renal failure.Kidney Int. 2001; 60: 2205-2214Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar A major concern in designing clinical trials is whether results in rodent models are translatable to man.10.Lieberthal W. Nigam S.K. Acute renal failure II. Experimental models of acute renal failure: imperfect but indispensable.Am J Physiol Renal Physiol. 2000; 278: F1-F12PubMed Google Scholar We therefore conducted these studies in large mammals in an attempt to better set the stage for human studies. In addition, there is increasing concern that ischemic acute renal failure leads to long-term fibrotic changes.11.Basile D.P. Donohoe D. Roethe K. Osborn J.L. Renal ischemic injury in permanent damage to peritubular capillaries and influence long-term function.Am J Physiol Renal Physiol. 2001; 281: F887-F899Crossref PubMed Scopus (573) Google Scholar, 12.Burne-Taney M.J. Yokota N. Rabb H. Persistant renal and extra renal immune changes after severe ischemic injury.Kidney Int. 2005; 67: 1002-1009Abstract Full Text Full Text PDF PubMed Scopus (83) Google Scholar Furthermore, findings in warm kidney IRI simulating native kidney injury may not be transferable to management of transplant patients who have a component of cold kidney IRI. The present study was designed to test the hypothesis that TBC-1269 reduced post-ischemic kidney damage in pig models of kidney IRI. The novel findings of this study include the following (1) TBC-1269 reduced kidney IRI in warm or cold ischemia in pigs; (2) the protective effect was associated with reduced CD4 and macrophage infiltration; and (3) long-term fibrosis was reduced by TBC-1269 treatment. The outcomes after warm ischemia and reperfusion differed markedly between experimental groups. Two animals from group warm ischemia (WI) 60 died on post-operative days 9 and 10. Survival was 100% in control group (Cont), uninephrectomized group (Unif), WI 45, WI TBC 45, and WI TBC 60. Renal functional data are shown in Figure 2a and b. During the first week, Ccr was reduced in all experimental groups (WI 45 to WI 60) when compared to control and sham-operated groups (Figure 2a). However, Ccr was significantly increased in WI TBC 45 when compared to WI 45, between weeks 2 and 16, and in WI TBC 60, when compared to WI 60 particularly after day 7. In group WI 45, Ccr paralleled Cont and sham-operated groups but remained at lower level from Cont. Ccr remained at 60% of Cont and Unif value in group WI 60. TBC-1269 also provided effective kidney protection after 24 h cold preservation (Figure 2b). However, no change was noted when TBC-1269 was only added to the preservation solution without administering at reperfusion. Two animals were excluded in CI24 group because of renal artery thrombosis. In the warm ischemia model, proteinuria was detected and peaked at day 3 and decreased thereafter to day 7. Progressive proteinuria developed again after approximately 7–8 weeks (Figure 3a). A significant difference was detected between WI 45 and WI 60 groups, and TBC-1269-treated groups, particularly after 7–8 weeks. In the cold ischemia model, proteinuria was more pronounced during the first week when compared to the warm ischemia model (Figure 3b). TBC-1269 also reduced proteinuria in cold-stored and autotransplanted groups. TBC-1269 when only added in the preservation solution had no influence in the outcome of proteinuria. TBC-1269 studied in animals submitted to the same Cont group conditions did not induce any change in the renal functions when compared to Cont group (Figure 3a and b). Histological analysis of biopsy samples from warm-ischemia kidneys revealed differences in the tissue injury after 40 min of reperfusion and of the day-7 post-reperfusion. Kidneys from WI 45 and WI 60 had increased damage when compared to WI TBC 45 and particularly WI TBC 60 (Table 1). The degree of initial acute proximal tubule cell injury appears closely related to the duration of warm ischemia, particularly after 60 min of WI. In the cold preservation and transplantation model, TBC-1269 was also effective in reducing renal tubular injury when compared to controls. TBC-1269 added in the preservation solution alone without reperfusion was not protective when compared to TBC CI 24 group that received the agent during preservation and reperfusion.Table 1Degree of proximal tubule cell lesions and mitochondria damage after different ischemia conditionsIschemic conditionsWarm ischemiaInjury type time reperfusionWI 45WI TBC 45WI 60WI TBC 60Brush border integrity 30–45 min3.5±0.23.4±0.22.6±0.2°3.1±0.2* Day 73.7±0.23.6±0.23.0±0.1°3.5±0.2*Cell detachment 30–45 min2.6±0.22.5±0.33.4±0.22.7±0,2* Day 71.5±0.21.4±0.22.7±±0.22.2±0.2*Ischemic conditionsCold ischemiaInjury type time reperfusionCI24TBCCI24TBCCI24ABrush border integrity 30–45 min2.5±0.23.6±0.23.5±0.2# Day 73.5±0.24.2±0.24.0±0.1#Cell detachment 30–45 min2.4±0.21.9±0.22.0±0.2# Day 71.7±0.21.3±0.21.2±0.2#The histological lesions from early (30–45 min) and day 7 after warm ischemia were evaluated and given as a semiquantitative score (Materials and Methods). Brush border integrity and tubular necrosis in light microscopy studies. Values are means±s.d. for all parameters analyzed.*P<0.05 WI 90 vs WI 60, °P<0.05 WI 60 vs WI 30 and WI 45, #P<0.05 TBC CI 24 and TBC CI 24A vs CI 24. Open table in a new tab The histological lesions from early (30–45 min) and day 7 after warm ischemia were evaluated and given as a semiquantitative score (Materials and Methods). Brush border integrity and tubular necrosis in light microscopy studies. Values are means±s.d. for all parameters analyzed. *P<0.05 WI 90 vs WI 60, °P<0.05 WI 60 vs WI 30 and WI 45, #P<0.05 TBC CI 24 and TBC CI 24A vs CI 24. Consistent with our previous studies in the pig model, the major inflammatory cell population detected by immunohistochemical long term studies after ischemia was CD4+ T lymphocytes. No infiltrating cells were detected in Cont and Unif (data not shown). In contrast, the number of CD4+ increased between weeks 1 and 2 after reperfusion in WI 45 and particularly WI 60 (Figure 4a). After week 2, CD4+ cells plateaued up to 16 weeks following reperfusion in groups WI 45 and WI 60. In WI 60 and WI 45 groups, monocyte/macrophage increased between weeks 1 and 2 and slightly decreased between weeks 2 and 4, then plateaued between weeks 4 and 16 (Figure 4b). Monocyte/macrophage number was significantly increased in WI 60 group when compared to WI 45 group. In WI kidneys treated with TBC-1269, CD4-positive cell number was significantly reduced. Monocyte/macrophage number was also different in WI TBC 45 and WI TBC 60 when compared to WI 45 and WI 60, respectively (Figure 5a). TBC-1269 was also efficient in cold ischemia model in reducing inflammatory cells (Figure 5b). TBC-1269 reduced CD4-positive cells and macrophages during the first week, and this beneficial effect remained detectable between weeks 4 and 16.Figure 5Influence of different ischemia models and administration of TBC-1269 on monocyte/macrophages within kidney tissue (4a and b). The number of positively stained cells per surface area was counted on biopsy samples from experimental groups. The values reported are the mean±s.d. from 8 to 12 separate determinations. #P<0.05, ##P<0.05 WI 45 vs WI TBC 45 and 0P<0.05, 000P<0.05 WI 60 vs WI TBC 60, **P<0.01 and ***P<0.001 CI 24 vs TBC CI 24, and TBC CI 24A.View Large Image Figure ViewerDownload (PPT) Evolution of interstitial fibrosis is presented in Figure 4. In our hands, fibrosis is correlated with WI, particularly after 60 min (Figure 6, lower panel, a–d). Percentage of interstitial fibrosis area is also associated with progression of renal dysfunction particularly in WI 60 group. Picrosirius score was significantly different at weeks 7–8 between WI 45 and WI 60 groups. These changes are accompanied by atrophy of the surrounding tubules (Figure 6, lower panel, e–h, upper panel). This difference in fibrosis increased at weeks 10–12 and more significantly by week 16. TBC-treated groups showed a significantly reduced fibrosis intensity particularly after 60 min of WI between weeks 8 to 16 (WI 60 vs WI TBC 60; Figure 6, lower panel) and weeks 10–12 and W 16 (WI 45 vs WI TBC 45; Figure 6). No difference was found at week 8 in WI 45 and WI TBC 45. Tubular atrophy scores were increased in WI 45 and WI 60 groups after week 8 when compared to TBC-1269-treated groups (Figure 6). In the cold ischemia model, TBC also reduced fibrosis (Figure 7, upper panel, a–c, lower panel) and tubular atrophy development (Figure 7, upper panel, d–f, lower panel).Figure 7Influence of cold ischemia on the onset of tubulointerstitial fibrosis and tubular atrophy score: the percentage of kidney areas displaying interstitial fibrosis stained with Picrosirius was measured over several weeks following cold ischemia within kidneys. *P<0.05 and **P<0.01 CI 24 vs TBC CI 24 and TBC CI 24A.View Large Image Figure ViewerDownload (PPT) IRI is the primary cause of acute renal failure in native kidney and can cause delayed graft function in transplanted kidneys. Ischemic injury to the kidney is associated with high morbidity and mortality and is a non-immunologic factor involved in chronic allograft nephropathy. In addition, recent report has demonstrated that patients with poor levels of early graft function are at an increased risk of death.13.Fernández-Fresnedo G. Rodrigo E. Escallada R. et al.Effect of early graft function on patient survival in renal transplantation.Transplant Proc. 2003; 35: 1653-1654Abstract Full Text Full Text PDF PubMed Scopus (5) Google Scholar The lack of suitable organs for transplantation from traditional sources has led clinicians to seek alternatives. Although related and unrelated live donation is being promoted as one option for increasing the donor pool, it is unlikely that this will in itself be able to completely bridge the gap. Non-heart beating and extended criteria organs, both typified by increased injury, can provide an alternative supply of organs, which should substantially increase the donor pool.14.Brook N.R. Waller J.R. Nicholson M.L. Nonheart-beating kidney donation: current practice and future developments.Kidney Int. 2003; 63: 1516-1529Abstract Full Text Full Text PDF PubMed Scopus (75) Google Scholar Consequently, therapeutics directed to improving the ability of the kidney to tolerate ischemic injury would have important implications. We found that selectin ligand binding with TBC-1269 significantly reduced post-ischemic renal dysfunction in pigs with different warm ischemia times, as well as in a cold ischemia model. Importantly, these findings on warm ischemia in a large mammal model confirm and extend earlier studies on rodents. Furthermore, we have evaluated a cold ischemia model as well, and demonstrated that administration in the perfusate is not adequate but that post-reperfusion dosing is crucial. Proteinuria occurred in the WI 45 and WI 60 groups, as well as CI 24 groups, and correlated with histologic injury and particularly tubulo-intersitital fibrosis. Proteinuria became prominent and progressive after week 8 in warm ischemia groups when compared to control and Unif. Interestingly, the protective effect of TBC-1269 was also prominent, long-term after warm ischemia. In cold preserved groups, TBC-1269-treated groups had reduced proteinuria and reduced fibrosis and tubular atrophy. We examined mononuclear leukocytes as potential mediators of late changes after IRI, and examined the impact of the selectin ligand blockade on these. TBC-1269 reduced monocyte/macrophage infiltration in WI TBC 45 and WI TBC 60 groups after weeks 6 and 8, respectively. We also evaluated the effects of TBC-1269 on CD4+ cells infiltration, which increased with WI time. TBC-1269 reduced CD4+ cell infiltration in both the WI model and the cold ischemia autotransplanted model. We cannot exclude the possibility that reduced macrophages and CD4+ cells were a result of decreased injury, rather than the cause of decreased injury. Recent studies have demonstrated that the in vivo effects of TBC-1269 may be mediated through E-selectin but do not appear to involve leukocyte rolling. TBC-1269 was shown to inhibit rolling of a murine cell line (32DC13) on murine P-selectin in vitro and has pronounced anti-inflammatory activity in mice when given before induction of thioglycollate-induced peritonitis.15.Hicks A.E.R. Abbitt K.B. Dodd P. et al.The anti-inflammatory effects of a selectin ligand mimetic TBC-1269, are not a result of competitive inhibition of leukocyte rolling in vivo.J Leukoc Biol. 2005; 77: 59-66PubMed Google Scholar In contrast, in this previous study, TBC-1269 failed to influence established leukocyte rolling in vivo at doses that match or exceed those with pronounced, anti-inflammatory activity. The authors hypothesized that TBC-1269 limits inflammation by a mechanism other than direct competition with cell-bound selectin ligands. An other recent study has demonstrated that TBC-1269 downregulated the expression of macrophage inflammatory protein (MIP)-1α, MIP-1β, MIP-2, and kupffer cells, as well as expression of Akt, mitogen-activated protein kinases (p44/42), and ribosomal S6 kinase.16.Toledo-Pereyra L.H. Lopez-Neblina F. Reuben J.S. et al.Selectin inhibition modulates Akt/MAPK signaling and chemokines expression after liver ischemia–reperfusion.J Invest Surg. 2004; 17: 303-313Crossref PubMed Scopus (32) Google Scholar Consequently, this selectin inhibition modulated the expression of Akt, mitogen-activated protein kinases (p44/42), and alterations in cellular signaling and chemokines. These alterations in cellular signaling and chemokine expression represent potential mechanisms or pathways of inflammatory response in ischemia-reperfusion. Mitogen-activated protein kinases have been the focus of a number of studies, as these compounds are involved in a number of important inflammatory cell signaling mechanisms.17.Lai E.W. Toledo-Pereyra L.H. Walsh J. et al.The role of MAP kinase in trauma and ischemia–reperfusion.J Invest Surg. 2004; 17: 45-53Crossref PubMed Scopus (27) Google Scholar Recent studies have further elucidated the role of mitogen-activated protein kinases in the inflammatory response, as a result of trauma and/or IRI. Akt pathway is also involved in IRI and functional recovery of damaged organs. It was recently shown that biosmanose was the most advanced synthetic pan-selectin antagonist and that TBC-1269 competitively inhibited, via steric/allosteric mechanisms, the binding of selectin function blocking antibody and was shown to bind all the three selectins to varying degrees.18.Beauharnois M.E. Lindquist K.C. Marathe D. et al.Affinity and kinetics of sialyl lewis-X and core-2 based oligosaccharides binding to L- and P-selectin.Biochemistry. 2005; 44: 9507-9519Crossref PubMed Scopus (64) Google Scholar, 19.Aydt E. Wolf G. Development of synthetic pan-selectin antagonists: a new treatment strategy for chronic inflammation in asthma.Pathobiology. 2003; 70: 297-301Crossref Scopus (37) Google Scholar Previous studies have demonstrated that renal, ischemic injury results in permanent damage to peritubular capillaries and influence long-term function.20.Basile D.P. Rarefaction of peritubular capillaries following ischemic acute renal failure: a potential factor predisposing to progressive nephropathy.Curr Opin Nephrol Hypertens. 2004; 13: 1-7Crossref PubMed Scopus (181) Google Scholar, 21.Bonventre J.V. Weinberg J.M. Recent advances in the pathophysiology of ischemic acute renal failure.J Am Soc Nephrol. 2003; 14: 2199-2219Crossref PubMed Scopus (635) Google Scholar Tubulo-interstitial fibrosis was significant in both warm and cold ischemia groups, long term. Picrosirius red-positive staining has been used for nearly 30 years and the use of computer-assisted quantification of fibrosis in the Sirius red-stained renal allograft biopsies appears to be a surrogate marker to predict long-term allograft function.22.Junqueira L. Bignoles G. Brentani R. Picro Sirius staining plus polarization microscopic, a specific method for collagen detection in tissue sections.Histochem J. 1979; 11: 447-455Crossref PubMed Scopus (1958) Google Scholar In our model, Picrosirius red staining was improved by the selectin ligand inhibition. These data sets the stage for a clinical trial applying selectin ligand blockade to improve short- and long-term outcomes of kidney IRI in both the native kidney and allograft. We have noted some difference between warm ischemia and cold preservation, particularly for proteinuria and fibrosis development. Inflammatory cell kinetics were also specific for cold and warm ischemia. A recent study demonstrated that WI triggers injury primarily to proximal tubular cells, whereas cold ischemia damages glomerular podocytes and peritubular endothelial cells in addition to proximal tubules.23.Yin M. Currin R.T. Peng X.X. et al.Different patterns of renal cell killing after warm and cold ischemia.Renal Failure. 2002; 24: 147-173Crossref PubMed Scopus (21) Google Scholar These data support proteinuria output in cold ischemia group. As previously reported, these results suggest that organ preservation and cold ischemia could have pathophysiology distinct from warm IRI, particularly after 60 min of WI.24.Kieran N.E. Rabb H. Immune response in kidney preservation and reperfusion injury.J Investig Med. 2004; 52: 310-314Crossref PubMed Google Scholar The fact that TBC-1269 does not seem to have any beneficial impact when added to the preservation solution could be linked to the change of the drug effect at 4°C. In addition, future studies are required to examine the molecular mechanisms of how selectin ligand blockade modulates kidney outcomes after IRI. Large white male pigs, 30–35 kg, 3–4 month old (INRA, Le Magneraud, Surgères, France) were prepared as previously described.25.Hauet T. Goujon J.M. Baumert H. et al.Polyethylene glycol reduces the inflammatory injury due to cold ischemia/reperfusion in autotransplanted pig kidneys.Kidney Int. 2002; 62: 654-667Abstract Full Text Full Text PDF PubMed Scopus (90) Google Scholar All animals were fed with a standard diet (Société Arrive, Saint Fulgent, France) and had free access to tap water. Rapid preoperative tranquilization was achieved by the rapid and atraumatic nasal administration of 0.2 mg/kg of midazolam (Laboratoire Roche, Neuilly-sur-Seine, France). The animals were then anesthetized with halothane (Laboratoire Belamont, Paris, France) and 100% oxygen. The surgical procedures were performed under sterile conditions. A 20-gauge plastic catheter (Becton Dickinson Vascular Access Inc., Sandy, UT, USA) was inserted into an ear vein. Atropine sulfate 10 μg/kg was given intravenously to reduce pharyngeal and tracheal secretion, and to prevent post-intubation bradycardia. A midline abdominal incision was made to expose both kidneys. Warm ischemia was induced in the pigs by clamping the left renal pedicle for 45 min (WI 45, n=10) and 60 min (WI 60, n=10) with a vascular clamp followed by a right nephrectomy. Two other experimental groups were studied: WI TBC 45 (n=10) and WI TBC 60 (n=10). In these two experimental groups, TBC-1269 was administered 10 min before ischemia, during warm ischemia, and 1 h after reperfusion. TBC was given at a concentration of 25 mg/kg based on a previous dosing study.26.Kogan T.P. Duprè B. Bui H. et al.Novel synthesis inhibitors selectin mediated cell adhesion: synthesis of 1,6-bis(3-(3 carboxy-methylphenyl)-4-(2-α-D-mannopyranosylloxy)phenyl)hexane-(TBC-1269).J Med Chem. 1998; 41: 1099-1111Crossref PubMed Scopus (134) Google Scholar In the cold ischemia model, the left kidney was removed and cold-flushed with University of Wisconsin solution and cold-stored at 4°C for 24 h. The kidneys were re-implanted and the contralateral kidney was removed. Groups of animals were divided as follows: CI TBC 24, n=10 (TBC-1269 was administered 10 min before cold-flush and 1 h after reperfusion), CI TBC 24A (CI TBC 24 with TBC-1269 added in UW solution before cold-flush but not reperfusion, n=10), and CI 24, n=10. These experimental groups were compared to uninephrectomy group without kidney ischemia (Unif; n=8) and control group (Cont; n=6, age-matched group). TBC-1269 was also studied in control group condition (Cont+TBC, n=6). All experiments were carried out in accordance with the Guidelines of the French Agricultural Office and the legislation governing animal studies. Any animal that died during the 16-week follow-up period was autopsied. All animals that died from causes other than renal failure, or developed renal artery, vein, or ureter problems were excluded. Endogenous creatinine clearance (Ccr, expressed in ml/min) and urinary protein were measured as previously described.25.Hauet T. Goujon J.M. Baumert H. et al.Polyethylene glycol reduces the inflammatory injury due to cold ischemia/reperfusion in autotransplanted pig kidneys.Kidney Int. 2002; 62: 654-667Abstract Full Text Full Text PDF PubMed Scopus (90) Google Scholar Endogenous creatinine clearance is suggested to be a reliable indicator for renal excretory function in newborn and growing piglets.27.Bauer R. Walter B. Zwiener U. Comparison between inulin clearance and endogenous creatinine in newborn normal weight and growth restricted newborn piglets.Exp Toxicol Pathol. 2000; 52: 367-372Crossref PubMed Scopus (13) Google Scholar, 28.Nielsen T.W. Maaske C.A. Booth N.H. Some comparative aspects of porcine renal function.in: Bustad L.K. McClellan R.O. Swine in Biomedical Resaerch. Frayn, Seattle, WA1996: 529-536Google Scholar Pigs were placed in a cage to allow specific 24 h urine collections and plasma samples collection. Plasma and urinary creatinine (Cr) was measured enzymatically using an automatic analyzer (Hitachi, Paris, France) and Ccr was calculated using the following formula: urinary flow rate × urinary Cr/plasma Cr. Urinary proteins was determined using a photometry method (Laboratoire Biorea, Talant, France). After warm ischemia and 40 min of reperfusion, at day 1, day 3, day 5, day 7, day 14 and 4–5, 10–12, and 16 weeks after surgery, biopsy tissue samples from the deep cortex–outer medulla region were performed. Samples were fixed with Dubosq-Brasil and 10% formalin in 0.01 mmol/l phosphate buffer (pH 7.42) and embedded in paraffin. Conventional stains were applied (hematoxylin and eosin, periodic acid-Schiff). Different histological parameters (brush border integrity and intratubular detachment) were studied. Histological lesions were expressed in percent of kidney samples using a previously described semiquantitative scale24.Kieran N.E. Rabb H. Immune response in kidney preservation and reperfusion injury.J Investig Med. 2004; 52: 310-314Crossref PubMed Google Scholar, 25.Hauet T. Goujon J.M. Baumert H. et al.Polyethylene glycol reduces the inflammatory injury due to cold ischemia/reperfusion in autotransplanted pig kidneys.Kidney Int. 2002; 62: 654-667Abstract Full Text Full Text PDF PubMed Scopus (90) Google Scholar: (0) no abnormality; (1) mild lesions affecting less than 10% of kidney samples; (2) lesions affecting 10–25% of kidney samples; (3) lesions affecting 25–50% of kidney samples; (4) lesions affecting 50–75%; (5) lesions affecting more than 75% of kidney samples. Tubulointerstitial injury was defined as inflammatory cell infiltrates and interstitial fibrosis. To estimate the level of tubulointerstitial fibrosis, tissue sections were also labeled with Picrosirius for collagen identification (collagen I and III) as previously described.29.Goujon J.M. Hauet T. Menet E. et al.Histological evaluation of proximal tubule cell injury in isolated perfused pig kidneys exposed to cold ischemia.J Surg Res. 1999; 82: 228-233Abstract Full Text PDF PubMed Scopus (55) Google Scholar, 30.Hauet T. Goujon J.M. Vandewalle A. et al.Trimetazidine reduces renal dysfunction by limiting the cold ischemia/reperfusion injury in autotransplanted pig kidneys.J Am Soc Nephrol. 2000; 11: 138-148PubMed Google Scholar The amount of interstitial fibrosis was determined in Picrosirius-stained sections by a semiquantitative imaging technique.31.Grimm P.C. Nickerson P. Gough J. et al.Computerized image analysis of Sirius Red-staining renal allograft biopsies as a surrogate marker to predict long-term allograft function.J Am Soc Nephrol. 2003; 14: 1662-1668Crossref PubMed Scopus (164) Google Scholar The percentage of Picrosirius-stained surface was determined on 10 different tissue sections viewed at × 100 magnifications in each experimental condition and expressed as percent of the total surface area examined. Tubular atrophy was semiquantitatively scored on a scale of 0 to 5+ by two pathologists blinded to the experimental conditions (0, normal; 1, small changes affecting 5–10%; 2, changes affecting 10–25% of specimen area; 3, changes affecting 25–50% of specimen area; 4, changes affecting 50–75% of specimen area; 5, changes affecting 75–100% of specimen area). Frozen and paraffin-embedded kidney biopsy sections (5 μm) from biopsies were processed for indirect immunohistochemistry using mouse monoclonal antibodies to detect macrophages (mouse antiporcine macrophage and monocyte, MCA1218, Serotec products, Oxford, UK) and CD4+ cells (mouse antiporcine CD4+, MCA1749, Serotec products). Sections were deparaffined, rehydrated, and heated in a pressure cooker containing citrate buffer (pH 6) to boiling point for 2 min. The sections were then cooled, rinsed in phosphate-buffered saline, and processed for indirect immunohistochemistry as previously described20.Basile D.P. Rarefaction of peritubular capillaries following ischemic acute renal failure: a potential factor predisposing to progressive nephropathy.Curr Opin Nephrol Hypertens. 2004; 13: 1-7Crossref PubMed Scopus (181) Google Scholar, 24.Kieran N.E. Rabb H. Immune response in kidney preservation and reperfusion injury.J Investig Med. 2004; 52: 310-314Crossref PubMed Google Scholar by using the antibodies listed in Table 1. All sections were examined under blinded conditions and photographed. The number of MCA1218- and CD4+-labeled cells per surface area (104/μm2) was counted on 10 different tissue sections for each of the experimental conditions as described.29.Goujon J.M. Hauet T. Menet E. et al.Histological evaluation of proximal tubule cell injury in isolated perfused pig kidneys exposed to cold ischemia.J Surg Res. 1999; 82: 228-233Abstract Full Text PDF PubMed Scopus (55) Google Scholar Values are reported as the mean±s.d. from (n) experiments. Statistical significance was assessed using the unpaired t-test and the Mann–Whitney U-test or by one-way analysis of variance using the Bonferroni t-test or the Newman–Keuls test for comparison of two or more than two groups. A P-value <0.05 was considered significant. This work was supported by grants from European Community (QLCK6-CT-2000-00064), from Région Poitou Charentes and from Centre Hospitalier et Universitaire de Poitiers, Banque Tarneaud and an NIH R01 to HR. We thank Goujon JM and Barriere M for their precious criticisms and Lochon D, Ribardiere F, Challeroux C, Boutin JL, Quellard N, and Fernandez B for their technical assistance. We thank Kurt Berens (TBC) who provided us with TBC-1269." @default.
• W2021983732 created "2016-06-24" @default.
• W2021983732 creator A5014171179 @default.
• W2021983732 creator A5025048132 @default.
• W2021983732 creator A5026312046 @default.
• W2021983732 creator A5026809269 @default.
• W2021983732 creator A5031850018 @default.
• W2021983732 creator A5035864437 @default.
• W2021983732 creator A5053012490 @default.
• W2021983732 creator A5077108704 @default.
• W2021983732 creator A5079664568 @default.
• W2021983732 date "2006-05-01" @default.
• W2021983732 modified "2023-10-18" @default.
• W2021983732 title "Protective role of selectin ligand inhibition in a large animal model of kidney ischemia–reperfusion injury" @default.
• W2021983732 cites W1801326471 @default.
• W2021983732 cites W1975361252 @default.
• W2021983732 cites W1979851813 @default.
• W2021983732 cites W1994205207 @default.
• W2021983732 cites W1994521388 @default.
• W2021983732 cites W2004662434 @default.
• W2021983732 cites W2015213347 @default.
• W2021983732 cites W2015781874 @default.
• W2021983732 cites W2032587462 @default.
• W2021983732 cites W2041689111 @default.
• W2021983732 cites W2046586570 @default.
• W2021983732 cites W2052927633 @default.
• W2021983732 cites W2055502077 @default.
• W2021983732 cites W2056065902 @default.
• W2021983732 cites W2056745009 @default.
• W2021983732 cites W2059652694 @default.
• W2021983732 cites W2092574818 @default.
• W2021983732 cites W2095134488 @default.
• W2021983732 cites W2097010504 @default.
• W2021983732 cites W2099675490 @default.
• W2021983732 cites W2101015806 @default.
• W2021983732 cites W2101538674 @default.
• W2021983732 cites W2128488710 @default.
• W2021983732 cites W2130492341 @default.
• W2021983732 cites W2157775555 @default.
• W2021983732 cites W2160800132 @default.
• W2021983732 cites W2328178163 @default.
• W2021983732 cites W4254671684 @default.
• W2021983732 doi "https://doi.org/10.1038/sj.ki.5000335" @default.
• W2021983732 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/16625150" @default.
• W2021983732 hasPublicationYear "2006" @default.
• W2021983732 type Work @default.
• W2021983732 sameAs 2021983732 @default.
• W2021983732 citedByCount "29" @default.
• W2021983732 countsByYear W20219837322012 @default.
• W2021983732 countsByYear W20219837322014 @default.
• W2021983732 countsByYear W20219837322016 @default.
• W2021983732 countsByYear W20219837322017 @default.
• W2021983732 countsByYear W20219837322018 @default.
• W2021983732 countsByYear W20219837322019 @default.
• W2021983732 countsByYear W20219837322020 @default.
• W2021983732 countsByYear W20219837322021 @default.
• W2021983732 countsByYear W20219837322022 @default.
• W2021983732 countsByYear W20219837322023 @default.
• W2021983732 crossrefType "journal-article" @default.
• W2021983732 hasAuthorship W2021983732A5014171179 @default.
• W2021983732 hasAuthorship W2021983732A5025048132 @default.
• W2021983732 hasAuthorship W2021983732A5026312046 @default.
• W2021983732 hasAuthorship W2021983732A5026809269 @default.
• W2021983732 hasAuthorship W2021983732A5031850018 @default.
• W2021983732 hasAuthorship W2021983732A5035864437 @default.
• W2021983732 hasAuthorship W2021983732A5053012490 @default.
• W2021983732 hasAuthorship W2021983732A5077108704 @default.
• W2021983732 hasAuthorship W2021983732A5079664568 @default.
• W2021983732 hasBestOaLocation W20219837321 @default.
• W2021983732 hasConcept C116569031 @default.
• W2021983732 hasConcept C126322002 @default.
• W2021983732 hasConcept C164705383 @default.
• W2021983732 hasConcept C170493617 @default.
• W2021983732 hasConcept C2778117688 @default.
• W2021983732 hasConcept C2780091579 @default.
• W2021983732 hasConcept C2780114680 @default.
• W2021983732 hasConcept C2780472472 @default.
• W2021983732 hasConcept C2780721119 @default.
• W2021983732 hasConcept C3018697912 @default.
• W2021983732 hasConcept C541997718 @default.
• W2021983732 hasConcept C56701166 @default.
• W2021983732 hasConcept C71924100 @default.
• W2021983732 hasConcept C89560881 @default.
• W2021983732 hasConcept C98274493 @default.
• W2021983732 hasConceptScore W2021983732C116569031 @default.
• W2021983732 hasConceptScore W2021983732C126322002 @default.
• W2021983732 hasConceptScore W2021983732C164705383 @default.
• W2021983732 hasConceptScore W2021983732C170493617 @default.
• W2021983732 hasConceptScore W2021983732C2778117688 @default.
• W2021983732 hasConceptScore W2021983732C2780091579 @default.
• W2021983732 hasConceptScore W2021983732C2780114680 @default.
• W2021983732 hasConceptScore W2021983732C2780472472 @default.
• W2021983732 hasConceptScore W2021983732C2780721119 @default.
• W2021983732 hasConceptScore W2021983732C3018697912 @default.
• W2021983732 hasConceptScore W2021983732C541997718 @default.
• W2021983732 hasConceptScore W2021983732C56701166 @default.
• W2021983732 hasConceptScore W2021983732C71924100 @default.
• W2021983732 hasConceptScore W2021983732C89560881 @default.

• next