Abstract
Angiogenesis is a multistep process requiring endothelial cell activation, migration, proliferation and tube formation. We recently reported that elevated secretion of interlukin 8 (IL8) by myotubes (MT) from subjects with Type-2 Diabetes (T2D) reduced angiogenesis by human umbilical vein endothelial cells (HUVEC) and human skeletal muscle explants. This lower vascularization was mediated through impaired activation of the phosphatidylinositol 3-kinase (PI3K)-pathway. We sought to investigate additional signaling elements that might mediate reduced angiogenesis. HUVEC were exposed to levels of IL8 equal to those secreted by MT from non-diabetic (ND) and T2D subjects and the involvement of components in the angiogenic response pathway examined. Cellular content of reactive oxygen species and Nitrate secretion were similar after treatment with [ND-IL8] and [T2D-IL8]. CXCR1 protein was down-regulated after treatment with [T2D-IL8] (p < 0.01 vs [ND-IL8] treatment); CXCR2 expression was unaltered. Addition of neutralizing antibodies against CXCR1 and CXCR2 to HUVEC treated with IL8 confirmed that CXCR1 alone mediated the angiogenic response to IL8. A key modulator of angiogenesis is matrix metalloproteinase-2 (MMP2). MMP2 secretion was higher after treatment with [ND-IL8] vs [T2D-IL8] (p < 0.01). MMP2 inhibition reduced tube formation to greater extent with [ND-IL8] than with [T2D-IL8] (p < 0.005). The PI3K-pathway inhibitor LY294002 reduced IL8-induced MMP2 release. IL8 regulation of MMP2 release was CXCR1 dependent, as anti-CXCR1 significantly reduced MMP2 release (p < 0.05). These results suggest that high levels of IL8 secreted by T2D MT trigger reduced capillarization via lower activation of a CXCR1-PI3K pathway, followed by impaired release and activity of MMP2.
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Data produced during this study are available from the corresponding author upon reasonable request. No specific materials were produced during this study.
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Acknowledgements
We wish to thank Leslie Carter for technical assistance.
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This work was supported by Merit Review Award No. I01CX00635 from the United States (U.S.) Department of Veterans Affairs Clinical Sciences Research and Development Service. The contents do not represent the views of the U.S. Department of Veterans Affairs or the United States Government.
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Robert R. Henry is deceased. This paper is dedicated to his memory.
- Robert R. Henry
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YAL researched data and wrote the manuscript. TPC researched data, contributed to discussion and reviewed/edited the manuscript. RRH contributed to discussion and reviewed/edited the manuscript. TPC is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.
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Levy, Y.A., Ciaraldi, T.P. & Henry, R.R. Impaired capillary tube formation induced by elevated secretion of IL8 involves altered signaling via the CXCR1/PI3K/MMP2 pathway. Mol Biol Rep 48, 601–610 (2021). https://doi.org/10.1007/s11033-020-06104-z
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DOI: https://doi.org/10.1007/s11033-020-06104-z