title: Limb ischemia-induced increase in permeability is mediated by leukocytes and leukotrienes.
abstract: This study tests the role of white blood cells (WBC) and leukotrienes in mediating the increased microvascular permeability following ischemia and reperfusion. Anesthetized dogs (n = 23) underwent 2 hours of hind limb ischemia induced by tourniquet inflation to 300 mmHg. In untreated animals (n = 7), tourniquet release led after 5 minutes to a rise in plasma thromboxane (Tx) B2 levels from 360 to 1702 pg/ml (p less than 0.05); after 2 hours, lymph TxB2 concentration had risen from 412 to 1598 pg/ml (p less than 0.05). There were decreases in circulating WBC from 11,766 to 6550/mm3 and platelets from 230 to 155 x 10(3)/mm3. During reperfusion, popliteal lymph flow (QL) increased from 0.07 to 0.24 ml/hour (p less than 0.05), while the lymph/plasma (L/P) protein ratio was unchanged from 0.39, changes consistent with increased microvascular permeability. WBC depletion (n = 7) to 302/mm3 by hydroxyurea or nitrogen mustard attentuated (p less than 0.05) the reperfusion induced rise in plasma TxB2 from 91 to 248 pg/ml and prevented the increase in lymph TxB2 concentration. Within 5 minutes of tourniquet release WBC counts further decreased to 191/mm3 (p less than 0.05) and platelets declined from 175 to 93 x 10(3)/mm3 (p less than 0.05). QL increased from 0.07 to 0.12 ml/hour (p less than 0.05), lower than untreated animals (p less than 0.05), and the L/P protein ratio declined from 0.49 to 0.37 (p less than 0.05), dilutional changes consistent with increased filtration pressure but not permeability to protein. Pretreatment with the lipoxygenase inhibitor diethylcarbamazine (DEC) (n = 8) prevented the reperfusion-induced increase in plasma and lymph TxB2 levels (p less than 0.05) and the fall in WBC counts (p less than 0.05), while platelet counts declined from 381 to 210 x 10(3)/mm3 (p less than 0.05). QL rose from 0.09 to 0.23 ml/hour (p less than 0.05) during reperfusion, and the L/P protein ratio of 0.3 remained unchanged, a value lower than in untreated dogs (p less than 0.05). In two animals of each group, vascular recruitment was induced by tourniquet inflation to 50 mmHg. This led to a high QL of 0.25 ml/hour and a low L/P ratio of 0.18. In untreated animals during reperfusion, QL further increased to 1.3 ml/hour, and L/P ratio rose to 0.44, documenting increased vascular permeability. In contrast, reperfusion in leukopenic or diethylcarbamazine (DEC)-treated dogs with vascular recruitment, was not associated with increases in QL or the L/P protein ratio.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Surgery, Brigham and Women's Hospital, Boston, MA 02115.
authors.forename: J M
authors.initials: JM
authors.lastname: Klausner
authors.forename: I S
authors.initials: IS
authors.lastname: Paterson
authors.forename: C R
authors.initials: CR
authors.lastname: Valeri
authors.forename: D
authors.initials: D
authors.lastname: Shepro
authors.forename: H B
authors.initials: HB
authors.lastname: Hechtman
mesh_terms: D015121:6-Ketoprostaglandin F1 alpha / Q000032:analysis / Q000097:blood; D000818:Animals; D002199:Capillary Permeability; D004049:Diethylcarbamazine / Q000494:pharmacology; D004285:Dogs; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000097:blood* / Q000503:physiopathology; D007958:Leukocyte Count; D007962:Leukocytes / Q000502:physiology*; D015289:Leukotrienes / Q000502:physiology*; D008196:Lymph / Q000032:analysis; D010976:Platelet Count; D015427:Reperfusion Injury / Q000097:blood / Q000503:physiopathology; D013929:Thromboxane B2 / Q000032:analysis / Q000097:blood
chemical_list: D015289:Leukotrienes; D013929:Thromboxane B2; D015121:6-Ketoprostaglandin F1 alpha; D004049:Diethylcarbamazine