title: Bacterial lipopolysaccharide-stimulated release of tumor necrosis factor-alpha from the isolated rat heart: the effect of aprotinin and forskolin.
abstract: Aprotinin has been reported to reduce plasma levels of inflammatory cytokines associated with cardiopulmonary bypass (CPB). Because CPB is also associated with elevated levels of bacterial lipopolysaccharide (LPS) and LPS stimulates release of inflammatory cytokines from the heart we tested the hypothesis that aprotinin would inhibit cardiac release of tumor necrosis factor-alpha (TNF) provoked by LPS. Isolated rat hearts were perfused Langendorf style. After 30 minutes of equilibration LPS (100 ng/mL) was infused for 60 minutes. Timed samples of coronary effluent were collected at 0, 30, 60, 90, 120, and 150 minutes after the initiation of LPS for the measurement of coronary flow and the determination of TNF and cyclic AMP. Other hearts were perfused with buffer containing aprotinin [137 kallikrein-inhibiting units (KIU)/mL or 250 KIU/mL] and then infused with LPS. An additional group received forskolin (10 microM) and LPS. In hearts perfused as controls with buffer alone no TNF was detected in the coronary effluent. In hearts perfused with LPS TNF was reliably detected in the coronary effluent at 60 minutes (606 +/- 450 pg/min) and increased with time to a level of 1792 +/- 650 pg/min at 150 minutes. The addition of aprotinin had no significant effect on LPS-stimulated TNF release. For instance in hearts perfused with 137 KIU/mL aprotinin LPS-stimulated release at 150 minutes was 2141 +/- 732 pg/min and in hearts perfused with 250 KIU/mL LPS-stimulated TNF release was 2049 +/- 789 pg/min. Forskolin administration was associated with release of cyclic AMP from the heart and completely inhibited LPS-stimulated TNF release. We conclude that LPS stimulated release of TNF from the heart. Adding aprotinin to the perfusion buffer in either high or low concentrations did not attenuate LPS-stimulated cytokine release. Elevating myocardial cyclic AMP with forskolin completely attenuated LPS-stimulated TNF release.
authors.affiliation: Department of Surgery, Mercer University School of Medicine and Medical Center of Central Georgia, Macon, Georgia 31201, USA.
authors.forename: William
authors.initials: W
authors.lastname: McKinney
authors.forename: Walter H
authors.initials: WH
authors.lastname: Newman
authors.forename: Jerry G
authors.initials: JG
authors.lastname: Webb
authors.forename: Manuel R
authors.initials: MR
authors.lastname: Castresana
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000276:immunology / Q000627:therapeutic use*; D002315:Cardiopulmonary Bypass / Q000009:adverse effects*; D005576:Colforsin / Q000276:immunology / Q000627:therapeutic use*; D000242:Cyclic AMP / Q000097:blood / Q000276:immunology; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D006321:Heart / Q000187:drug effects* / Q000378:metabolism*; D066298:In Vitro Techniques; D007249:Inflammation / Q000097:blood / Q000188:drug therapy* / Q000276:immunology; D008070:Lipopolysaccharides / Q000009:adverse effects*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D005576:Colforsin; D007611:Aprotinin; D000242:Cyclic AMP