title: Effects of acute global venous obstruction and unfractionated heparin on muscle cytokine synthesis.
abstract: Phlegmasia cerulea dolens is a devastating complication of massive deep venous thrombosis, which is clinically characterized by massive lower extremity tissue edema and subsequent arterial insufficiency. These experiments evaluated the local tissue effects of acute global venous obstruction combined with partial arterial ischemia. Experiments were performed to assess the effects of heparin on the cytokine response to simultaneous venous and partial arterial obstruction. Murine hind limbs were subjected to conditions of unilateral venous occlusion and partial tourniquet limb ischemia, which was confirmed by laser Doppler imaging (LDI). Mice underwent either hind limb venous obstruction with intravenous unfractionated heparin (200IU/kg) or intravenous saline 5min before venous occlusion. Sham-treated mice were subjected to anesthesia alone without venous occlusion. After 3hr, the mice were killed and tissue was harvested for measurement of edema (wet to dry weight ratio, W/D), muscle viability, indices of local thrombosis (thrombin-antithrombin complex [TAT]), and cytokine analysis for growth-related oncogene-1 (GRO-1) and interleukin-6 (IL-6, protein via enzyme-linked immunoassay and mRNA via reverse transcriptase polymerase chain reaction). Bleeding time and volume were documented in saline- and heparin-treated mice to confirm systemic anticoagulation. Administration of intravenous heparin resulted in a marked increase in bleeding time and volume. LDI confirmed venous obstruction and ongoing arterial inflow. Venous obstruction resulted in severe visible edema that correlated with a significantly higher W/D ratio but was not associated with a significant decrease in muscle viability. GRO-1 and IL-6 protein and mRNA levels were significantly elevated in the venous occlusion group compared to sham. Heparin therapy significantly decreased TAT3 levels but did not alter the profile of GRO-1 or IL-6 protein levels seen with venous occlusion. Venous occlusion with partial ischemia induces a unique and potent local cytokine expression. Heparin therapy did not ameliorate the cytokine response. These data indicate that heparin therapy does not modulate the cytokine response to venous obstruction.
authors.affiliation: Division of Vascular and Endovascular Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: Hong T
authors.initials: HT
authors.lastname: Hua
authors.forename: Hassan
authors.initials: H
authors.lastname: Albadawi
authors.forename: Fateh
authors.initials: F
authors.lastname: Entabi
authors.forename: Mark F
authors.initials: MF
authors.lastname: Conrad
authors.forename: Michael C
authors.initials: MC
authors.lastname: Stoner
authors.forename: Stuart
authors.initials: S
authors.lastname: Houser
authors.forename: Michael T
authors.initials: MT
authors.lastname: Watkins
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000008:administration & dosage; D000990:Antithrombin III / Q000378:metabolism; D054360:Chemokine CXCL1 / Q000096:biosynthesis; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004195:Disease Models, Animal; D004487:Edema / Q000097:blood / Q000188:drug therapy / Q000276:immunology* / Q000503:physiopathology; D006493:Heparin / Q000008:administration & dosage; D006614:Hindlimb; D007275:Injections, Intravenous; D015850:Interleukin-6 / Q000096:biosynthesis; D007511:Ischemia / Q000097:blood / Q000188:drug therapy / Q000276:immunology* / Q000503:physiopathology; D017078:Laser-Doppler Flowmetry; D051379:Mice; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000276:immunology*; D010447:Peptide Hydrolases / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D012039:Regional Blood Flow; D013924:Thrombophlebitis / Q000097:blood / Q000188:drug therapy / Q000276:immunology* / Q000503:physiopathology; D014111:Tourniquets; D014689:Venous Insufficiency / Q000097:blood / Q000188:drug therapy / Q000276:immunology* / Q000503:physiopathology
chemical_list: D000925:Anticoagulants; D054360:Chemokine CXCL1; C475895:Cxcl1 protein, mouse; D016207:Cytokines; D015850:Interleukin-6; D012333:RNA, Messenger; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D006493:Heparin; D010447:Peptide Hydrolases