title: Transfection with a dominant-negative inhibitor of monocyte chemoattractant protein-1 gene improves cardiac function after 6 hours of cold preservation.
abstract: BACKGROUND
  Monocyte chemoattractant protein-1 (MCP-1), a potent chemotactic factor for monocytes, is induced during ischemia-reperfusion. As monocytes might play an important causative role in reperfusion injury, we investigated if inhibition of monocyte activation could attenuate ischemia-reperfusion injury and thereby improve cardiac preservation. To inhibit monocyte activation, we transfected a dominant-negative inhibitor of MCP-1 (7ND) gene in an animal model.
  
  
  RESULTS
  We used an isolated rabbit heart preparation perfused with support-rabbit blood and transfected 7ND genes to skeletal muscle of the support rabbits (n=7) using electroporation technique; causing an elevation of serum 7ND level to 20+/-7 pg/mL at 5 days after transfection. Animals receiving empty plasmid served as controls (n=7). Five days after transfection, hearts from other rabbits were excised, stored in UW solution for 6hours, and perfused with blood from transfected support rabbits. The 7ND group showed better cardiac output (128.7+/-17.9 versus 81.6+/-19.8 mL/min; P<0.01), lower serum CK-MB levels (5.0+/-1.8 versus 11.1+/-2.9 ng/mL; P<0.01), lower serum IL-1beta levels (257.2+/-23.2 versus 311.2+/-37.4pg/mL; P<0.05), and lower serum TNF-alpha levels (19.0+/-8.4 versus 35.1+/-13.0pg/mL; P<0.05). The numbers of infiltrating cells in myocardium were significantly reduced in the 7ND group.
  
  
  CONCLUSIONS
  Inhibition of MCP-1 with 7ND gene transfection reduced cytokine activation, attenuated myocardial damage, and improved cardiac function after 6 hours of preservation. These results show that MCP-1 plays an important role in ischemia-reperfusion injury.
authors.affiliation: Department of Cardiovascular Surgery, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Kajihara
authors.forename: Shigeki
authors.initials: S
authors.lastname: Morita
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nishida
authors.forename: Hideki
authors.initials: H
authors.lastname: Tatewaki
authors.forename: Masataka
authors.initials: M
authors.lastname: Eto
authors.forename: Kensuke
authors.initials: K
authors.lastname: Egashira
authors.forename: Hisataka
authors.initials: H
authors.lastname: Yasui
mesh_terms: D000818:Animals; D002465:Cell Movement; D018932:Chemokine CCL2 / Q000097:blood / Q000235:genetics*; D003080:Cold Temperature; D003402:Creatine Kinase / Q000097:blood; D052279:Creatine Kinase, MB Form; D006321:Heart / Q000502:physiology*; D007375:Interleukin-1 / Q000097:blood; D007527:Isoenzymes / Q000097:blood; D009000:Monocytes / Q000502:physiology; D009154:Mutation; D015428:Myocardial Reperfusion Injury / Q000473:pathology / Q000628:therapy; D009206:Myocardium / Q000473:pathology; D009926:Organ Preservation; D011817:Rabbits; D013997:Time Factors; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis; D016277:Ventricular Function, Left
chemical_list: D018932:Chemokine CCL2; D007375:Interleukin-1; D007527:Isoenzymes; D014409:Tumor Necrosis Factor-alpha; D003402:Creatine Kinase; D052279:Creatine Kinase, MB Form