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ATHEROSCLEROSIS AND INFLAMMATION
Baltaeva Yu. Yu.
*
, Research Assistant, Fergana Branch of The Tashkent Medical
Academy.
Prof. Hans A.R.Bluyssen, Department of Human Molecular Genetics, Adam Mickiewicz
University in Poznan, Poland.
Atherosclerosis, the morphological correlates of vascular disease, is characterized by early
endonthelial dysfunction, vascular inflammation together with build-up of lipids, cholesterol, calcium
and cellular debris within the intima of the vessel wall. This build-up leads to the formation of advanced
atherosclerotic plaque
(Hans A.R.Bluyssen,et al., 2012).
Despite the fact that better treatments have
relieved the number of deaths from atherosclerosis-related diseases, and have improved the quality of
life for people who have these diseases,
atherosclerosis remains the underlying cause of about 50% of all death in westernized society.
According to the data given by WHO (the World Health Organization), an estimated 17.9 million people
died from Cardiovascular Diseases (CVDs) in 2016, representing 31% of all global deaths. Of these
deaths, 85% are due to heart attack and stroke. Over three quarters of CVD deaths take place in low-
and middle-income countries, including Uzbekistan. Out of the 17 million premature deaths (under the
age of 70) due to noncommunicable diseases in 2015, 82% are in low- and middle-income countries,
and 37% are caused by CVDs. Despite the many novel insights from a hematologic, genetic, and
pharmacological research, currently available treatments are still not very effective and atherosclerosis
remains a common health problem and is still a major burden on humanity. This means that the search
for new therapeutical agents is necessary.
According to above mentioned,
the purpose
of the research is to identification of potential
novel atherosclerosis biomarkers.
Materials and methods.
Real-time Polymerase Chain Reaction (PCR) was used in order to
analyze and compare the expression of selected target genes. The expression levels were compared with
present RNA-sequencing data per each gene and each treatment condition. Human Microvascular
Endothelial Cells (HMEC) were provided by the Center for Disease Control and Prevention (Atlanta,
GA) and cultured in MCDB-131 medium(IITD PAN, Wroclaw, Poland) containing 10% of fetal bovine
serum (FBS) (Gibco, Thermo Fisher), 100 U/ml penicillin, 100 pg/ml streptomycin, 0.01 pg/ml EGF,
0.05 pM hydrocortisone and 2 mM L- glutamine. At least 12h before the experiment, full medium was
exchanged for serum starved- medium (containing 1% FBS instead of 10%). HMECs were treated with
25 U/ml of murine interferon-y (IFNy) alone for 8 hours and IFNy (purchased from Merck) treatment
was followed separately by treatment with 50 U/ml of Lipopolysaccharides (LPS)( provided by Sigma-
Aldrich) for additional 4 hours and at the end LPS alone for 4 hours to induce signal integration pathway
between IFNs and toll-like receptors (TLRs). Total DNA was isolated using GeneMATRIX Universal
DNA Purification Kit (EURx, Gdansk, Poland). The expression levels were compared to reference
genes Actin (Actb) for both Vascular smooth muscle cells (VSMCs) and human microvascular
epithelial cells (HMEC).
Real-time PCR analyses were compared for both HMECs and SMCs and certain treatment
conditions were considered as an important factor. All of the conducted PCR results were evaluated
and discussed in order to choose the most significant expression levels and genes. Among the genes
have selected, Cxcl10 showed significant up-regulation for the patient material samples from ischemic
heart disease in comparison to control subjects. This gene is expressed in both human and mouse
atherosclerotic plaques. Cxcl10 plays a fundamental role as an anti-inflammatory factor and induces
foam cell formation. To evaluate this gene in further studies, bigger group of samples are needed with
the different population profiles.
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