Related Experiment Video
Updated: Jul 18, 2026
Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions
Published on: August 8, 2012
Cardiotrophin-1 induces interleukin-6 synthesis in human umbilical vein endothelial cells
Michael Fritzenwanger1, Katharina Meusel, Martin Foerster
1Department of Internal Medicine I, Division of Cardiology, Friedrich-Schiller-University Jena, Erlanger Allee 101, 07740 Jena, Germany. Michael.Fritzenwanger@med.uni-jena.de
Insights
Cardiotrophin-1 (CT-1) increases interleukin-6 (IL-6) in human endothelial cells via JAK2, STAT3, p38, and NFkappaB pathways. This finding suggests CT-1 may contribute to elevated IL-6 in chronic heart failure (CHF).
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Cellular Signaling
Background:
- Chronic heart failure (CHF) is associated with elevated proinflammatory cytokines, including interleukin-6 (IL-6).
- Cardiotrophin-1 (CT-1), a member of the IL-6 superfamily, is also increased in CHF patients.
- The role of CT-1 in inducing IL-6 in endothelial cells and its underlying mechanisms in CHF remain to be fully elucidated.
Purpose of the Study:
- To investigate whether CT-1 induces IL-6 expression in human umbilical vein endothelial cells (HUVEC).
- To characterize the signaling pathway involved in CT-1-mediated IL-6 induction.
- To explore the potential contribution of CT-1 to elevated IL-6 levels in CHF.
Main Methods:
- HUVEC were stimulated with varying concentrations and durations of CT-1.
- IL-6 mRNA levels were quantified using real-time PCR and RT-PCR.
- IL-6 protein concentrations in the supernatant were measured by ELISA.
- Specific pathway inhibitors (AG490, SB203580, piceatannol, parthenolide, cycloheximide, wortmannin, PD98059) were used to determine the underlying signaling cascade.
Main Results:
- CT-1 significantly increased IL-6 mRNA and protein expression in HUVEC in a concentration- and time-dependent manner.
- Maximal induction of IL-6 mRNA was observed after 6 hours, and protein after 24 hours.
- The CT-1-induced IL-6 production was inhibited by AG490 (JAK2), SB203580 (p38 MAPK), piceatannol (STAT3), parthenolide (NFkappaB), and cycloheximide (protein synthesis).
- Wortmannin (PI3K) and PD98059 (MEK1/2) did not affect CT-1-induced IL-6 expression.
Conclusions:
- CT-1 induces IL-6 mRNA and protein expression in HUVEC through a pathway involving JAK2, STAT3, p38 MAPK, and NFkappaB activation.
- Protein synthesis is required for CT-1-induced IL-6 expression, and IL-6 is not stored intracellularly.
- CT-1 may partially contribute to increased IL-6 plasma concentrations in CHF.
- Targeting the CT-1 pathway could represent a novel therapeutic strategy for CHF.
Abstract:
In patients with chronic heart failure (CHF) increased plasma concentrations of proinflammatory cytokines are found. For example, the plasma interleukin-6 (IL-6) concentration correlates with disease severity. Beside IL-6 cardiotrophin-1 (CT-1), a member of the IL-6 superfamily, is also increased in CHF. We examined whether CT-1 is able to induce IL-6 in human umbilical vein endothelial cells (HUVEC) and characterised the underlying pathway. IL-6 mRNA was determined by real-time PCR and by RT-PCR in HUVEC which were stimulated with different CT-1 concentrations and for different time periods. IL-6 concentration in the supernatant was determined by ELISA. For the pathway determination following inhibitors were used: piceatannol (signal transducer and activation of transcription (STAT)3 phosphorylation), wortmannin (phosphatiylinositol 3-kinase (PI3K)), SB203580 (p38 mitogen-activated protein kinase (MAPK)), AG490 (Janus kinase (JAK)2), PD98059 (mitogen-activated protein kinase kinase (MEK) 1/2), parthenolide (nuclear factor kappaB) and cycloheximide (protein biosynthesis). CT-1 caused a concentration- and time-dependent increase in IL-6 mRNA in HUVEC with a maximal induction seen after 6 h (2-fold compared to control) with 100 ng/ml CT-1. In the supernatant of HUVEC a concentration- and time-dependent increase of IL-6 protein was found. A maximum effect with 100 ng/ml CT-1 was found after 24 h (11-fold compared to control). AG490, SB203580, piceatannol, parthenolide and cycloheximide inhibit CT-1 induced IL-6 mRNA and protein expression whereas wortmannin and PD98059 did not inhibit IL-6 expression. CT-1 induced both IL-6 mRNA and protein in a concentration- and time-dependent manner in HUVEC. The underlying pathway includes activation of JAK2, STAT3, p38 and NFkappaB. CT-1 induced IL-6 expression and requires protein synthesis and IL-6 is not stored intracellularly. We speculate that in CHF CT-1 might be in part responsible for increased IL-6 plasma concentrations. Modulation of the CT-1 pathway may be a further strategy in CHF treatment.
