EXPRESSION OF PRO-INFLAMMATORY CYTOKINES IN PRIMARY HUMAN CORONARY ARTERY AND INTERNAL THORACIC ARTERY ENDOTHELIAL CELLS

Highlights

• Primary arterial endothelial cells show an expression of ≈ 40 cytokines, including ≈ 25 molecules overexpressed ≥5-fold at pro-inflammatory endothelial dysfunction.
• Despite primary arterial endothelial cells expressed a variety of cell adhesion molecules, only ≈ 5 of them are overexpressed ≥5-fold at pro-inflammatory endothelial dysfunction.
• Primary human coronary artery endothelial cells exhibit higher basal expression of pro-inflammatory cytokines in comparison with internal thoracic artery endothelial cells.

Aim. To compare expression of the genes encoding pro-inflammatory cytokines and cell adhesion molecules in primary human coronary artery endothelial cells (HCAEC) and human internal thoracic artery endothelial cells (HITAEC) for the assessment of pro-inflammatory activation in the ECs isolated from atherosusceptible and atheroresistant arteries.

Methods. Following the dot blotting profiling, we quantified the expression of the genes encoding pro-inflammatory cytokines (MIF, IL6, CXCL8, CCL2, CCL5, CCL20, CSF2, CSF3, CXCL1, CXCL5, CXCL10, PTX3) and cell adhesion molecules (VCAM1, ICAM1, SELE, SELP) by RT-qPCR. We further performed an extended analysis of gene expression by interrogating three RNA sequencing datasets.

Results. We found ≈ 40 pro-inflammatory cytokines expressed in HCAEC and HITAEC. Baseline expression with TPM_Ctrl > 2.5 was detected for 24 genes: MIF, CCL2, PTX3, IL32, CXCL1, TGFB1, LTB, CXCL8, CSF1, CCL14, TGFB2, IL33, CXCL16, CXCL2, IL1A, CSF3, IL6, IL17D, CXCL3, CXCL6, IL12A, CXCL5, TGFB3, and CXCL12. In addition, 24 genes (CSF2, CCL5, CCL20, CXCL5, CXCL8, CXCL3, CXCL11, IL1A, CXCL6, IFNE, CCL16, CXCL2, LTB, LTA, IL23A, CSF1, CXCL1, CXCL10, CSF3, IL6, IL32, TGFB1, CCL2, and IL7) had fold change ≥ 1.50 in dysfunctional ECs as compared with the control ECs. Three genes encoding cell adhesion molecules (VCAM1, ICAM1, and SELE) had fold change > 2 and TPM_{Ctr l}> 2.5. In comparison with HITAEC, HCAEC had higher expression of pro-inflammatory genes (MIF, IL6, CCL5, CSF3, CXCL1, and SELP) indicating higher pro-inflammatory status.

Conclusion. HCAEC have higher expression of pro-inflammatory genes comparing to HITAEC, in concert with the higher atherosusceptibility of CA.

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