RepOD

COVID-19 is multisystemic with varying degrees of severity. Unfortunately, despite intensive research, the molecular changes caused by Sars-CoV-2 still remain unclear. Mechanisms affected by virus infection include endothelial dysfunction and angiogenesis. Similarly, the vaccines developed so far affect the process of angiogenesis, contributing to the development of undesirable effects on the part of the cardiovascular system. Therefore, we aimed to detect the impact of the SARS-CoV-2 infection and Pfizer Comirnaty vaccine on the molecular aspect of angiogenesis.

HIF-1α (hypoxia-inducible factor 1 subunit alpha), VEGFA (vascular endothelial growth factor A), MMP-2 (matrix metalloproteinase 2), MMP-7 (matrix metalloproteinase 7), MMP-9 (matrix metalloproteinase 9), TIMP1 (TIMP metallopeptidase inhibitor 1) and ADAMTS1 (ADAM metallopeptidase with thrombospondin type 1 motif 1) genes expression was determined by real-time PCR using species-specific TaqMan Gene Expression Assay (HIF-1α-assay ID: Hs00936368_m1, VEGFA-assay ID: Hs00900055_m1, MMP-2-assay ID: Hs01548727_m1, MMP-7-assay ID: Hs01042796_m1, MMP-9-assay ID: Hs00957562_m1, TIMP1-assay ID: Hs01092511_m1 and ADAMTS1 (assay ID: Hs00199608_m1), and 18S as reference gene – assay ID Hs99999901_s1; Thermo Fisher Scientific, Waltham, MA, USA) and RT PCR Mix Probe (A&A Biotechnology, Gdynia, Poland) according to the manufacturer’s instructions on CFX96™ Re-al-Time PCR Detection System Thermal Cycler (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Before real-time PCR, cDNA was synthesised using total RNA and Applied Biosystems High-Capacity cDNA Reverse Transcription Kit (Foster City, CA, USA) according to the manufacturer’s protocols. Finally, the level of mRNA expression was measured about that of the reference gene, which was 18S ribosomal RNA as internal mRNA control and relative mRNA expression levels were calculated using the 2^−ΔCt method. (This study included 33 COVID-19 convalescents having a positive test result for SARS-CoV-2 infection, as indicated by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) of nasopharyngeal swab samples, 35 healthy people vaccinated with one dose of BNT162b2, 19 convalescent (with a confirmed status by test) vaccinated with one dose of BNT162b2)).