title: Cardiotrophin-1 induces monocyte chemoattractant protein-1 synthesis in human umbilical vein endothelial cells.
abstract: In chronic heart failure (CHF) cardiotrophin-1 (CT-1) and monocyte chemoattractant protein-1 (MCP-1) plasma concentrations are elevated. CT-1 is a cytokine of the interleukin-6 (IL-6) superfamily. Most members of the IL-6 family are able to activate human umbilical vein endothelial cells (HUVEC) but so far there are no data which demonstrate that CT-1 can activate HUVEC. Because MCP-1-as a marker of endothelial activation-is elevated in CHF we examined whether CT-1 will induce MCP-1 production in HUVEC. MCP-1 mRNA levels were determined by real time PCR, RT-PCR and northern blot analysis and MCP-1 protein concentrations in the supernatant by ELISA. Signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (pSTAT3) were investigated by western blot analysis. Incubation of HUVEC with different CT-1 concentrations for various time periods induced time and concentration dependent MCP-1 mRNA. Maximal MCP-1 mRNA was reached after 6h. After 24h CT-1 caused a significant induction of MCP-1 protein in the supernatant compared to control. CT-1 induced concentration dependent phosphorylation of STAT3 without any change in total-STAT3 concentration. Piceatannol-a specific blocker of STAT3 phosphorylation-inhibited CT-1 induced MCP-1 induction completely. AG490-a blocker of the JAK2 pathway-was also able to inhibit CT-1 induced MCP-1 upregulation, indicating that the JAK2 pathway is also necessary for MCP-1 induction. Parthenolide-a blocker of NFkappaB-inhibited CT-1 induced MCP-1 expression, completely. Our data show that CT-1 induces in a concentration and time dependent manner MCP-1 mRNA and protein in HUVEC. STAT3 phosphorylation, the activation of JAK2 and NF-kappaB are involved in this pathway. In CHF, CT-1 may be able to induce MCP-1 which might be responsible for progression of heart failure either by recruiting inflammatory cells within the myocardium or by a direct modulation of myocyte function.
authors.affiliation: Department of Internal Medicine I, Division of Cardiology, Friedrich-Schiller-University Jena, Erlanger Allee 101, 07740 Jena, Germany. michael.fritzenwanger@med.uni-jena.de
authors.forename: Michael
authors.initials: M
authors.lastname: Fritzenwanger
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Kuethe
authors.forename: Daniela
authors.initials: D
authors.lastname: Haase
authors.forename: Enrico
authors.initials: E
authors.lastname: Jandt
authors.forename: Hans-R
authors.initials: HR
authors.lastname: Figulla
mesh_terms: D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism*; D016207:Cytokines / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism / Q000502:physiology; D013997:Time Factors; D014471:Umbilical Veins / Q000378:metabolism*
chemical_list: D018932:Chemokine CCL2; D016207:Cytokines; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; C092073:cardiotrophin 1