Hyperexpression of TLR2 and TLR4 in patients with ischemic stroke in acute period of the disease

Pathogenesis of ischemic stroke is actively involved in the system of innate immunity. Under conditions of cerebral ischemia, a number of biologically active substances are released that interact with innate immunity receptors, in particular TLR2 and TLR4, which exacerbate inflammation in brain tissue. Identification of predictor markers at the level of the innate immunity system may foresee the clinical course of ischemic stroke and ensure timely treatment. Our objective was to study expression of TLR2 and TLR4 receptors in peripheral blood leukocytes in patients with ischemic stroke in the dynamics of the disease. 27 people were included in the study. The main group consisted of patients with ischemic stroke of varying severity (n = 19). Patients of the main group were divided into two subgroups: with an NIHSS index value of < 10 (n = 10) and > 10 (n = 9). The control group included healthy donors with no history of acute and chronic inflammatory diseases (n = 8). Peripheral blood leukocytes were used as the test material. To determine expression of the TLR2 and TLR4 genes, RT-PCR in real time was used. Surface expression of TLRs was determined by flow cytometry. A study of the TLR2 and TLR4 gene expression showed that on the 1st, 3rd and 7th day post-stroke, the TLR4 gene expression in patients was significantly increased, when compared to the control group (p < 0.01), whereas TLR2 gene expression on the 3rd day of the disease was not statistically different from the control group. A study of surface expression of receptors showed that the average TLR2 fluorescence intensity on the patients’ peripheral blood monocytes was significantly increased on the 1st and 3rd day of disease when compared to the control group. The surface expression of TLR4 on monocytes has a statistically significant increase only on day 7. Assessment of surface expression of TLRs in subgroups with different severity values by NIHSS showed that patients with a NIHSS index > 10 had a significantly higher level of surface of TLR2 expression over the observation period, while the largest difference in TLR4 expression in the subgroups was observed on the 1st day of the disease (p < 0.05). Patients with ischemic stroke showed an increase in TLR2 and TLR4 expression at the gene and protein level, compared to healthy donors. These indices can be considered possible predictors for clinical prognosis of ischemic stroke.

Download Full-text

Expression of DROSHA and DICER genes in peripheral blood leukocytes in lung sarcoidosis

Terapevticheskii arkhiv ◽

10.26442/terarkh201890321-24 ◽

2018 ◽

Vol 90 (3) ◽

pp. 21-24

Author(s):

I E Malysheva ◽

O V Balan ◽

E L Tikhonovich ◽

T O Volkova

Keyword(s):

Gene Expression ◽

Polymerase Chain Reaction ◽

Real Time ◽

Peripheral Blood ◽

Messenger Rna ◽

Peripheral Blood Leukocytes ◽

Chain Reaction ◽

Blood Leukocytes ◽

Healthy Donors ◽

Polymerase Chain

Aim. To study the expression level of the genes DROSHA and DICER in peripheral blood leukocytes (PBL) of patients with sarcoidosis of the lungs Materials and methods. The study included 32 patients diagnosed with persistent lung sarcoidosis (mean age 41.56±1.27 years) and 36 healthy donors (control; mean age 42.79±1.95 years). The level of expression of messenger RNA (mRNA) of the genes DROSHA and DICER were determined in PBL of healthy donors and patients with sarcoidosis of the lung by polymerase chain reaction in real time. Results. As a result of the conducted researches it is established that the level of drosha gene expression in PBL patients with sarcoidosis of lungs is significantly reduced in comparison with the control (p

Download Full-text

Association of FOXP3 gene -3279 C>A polymorphism with the risk of pulmonary sarcoidosis

Terapevticheskii arkhiv ◽

10.17116/terarkh2017891264-67 ◽

2017 ◽

Vol 89 (12) ◽

pp. 64-67 ◽

Cited By ~ 1

Author(s):

I E Malysheva ◽

L V Topchieva ◽

E L Tikhonovich ◽

I V Kurbatova ◽

O V Balan

Keyword(s):

Peripheral Blood ◽

Polymorphic Marker ◽

Pulmonary Sarcoidosis ◽

Control Group ◽

Peripheral Blood Leukocytes ◽

Blood Leukocytes ◽

Foxp3 Gene ◽

Healthy Donors ◽

Gene Transcripts ◽

The Republic

Aim. To investigate the association of the polymorphic marker -3279 C>A of the FOXP3 gene with the risk of pulmonary sarcoidosis (PS) and to estimate the transcription level of this gene in the carriers of different genotypes of this polymorphic marker. Subjects and methods. The investigation included 99 patients of Russian ethnicity (mean age, 45.41±1.31 years) living in the Republic of Karelia, who were diagnosed with persistent PS, and 116 healthy donors (mean age, 42.06±1.30 years) in the control group. The alleles and genotypes of the polymorphic marker -3279 C>A of the FOXP3 gene were identified using polymerase chain reaction (PCR)-restriction fragment length polymorphism. The number of transcripts of the studied gene in the peripheral blood leukocytes of healthy donors and PS patients was determined with real-time PCR. Results. The control group and the PS patient one had no statistically significant differences in the distribution of the frequencies of alleles and genotypes by the polymorphic marker –308G>A of the FOXP3 gene (p > 0.05). The number of FOXP3 gene transcripts was not statistically significantly different in the peripheral blood leukocytes of patients with PS and control individuals. No statistically significant differences were observed in the mRNA expression levels in the above-mentioned gene in the carriers of different genotypes by the polymorphic marker -3279 C>A of the FOXP3 gene in all examined groups. Conclusion. The polymorphic marker -3279 C>A of the FOXP3 gene is unassociated with the risk of PS.

Download Full-text

Correlation of Adrenomedullin gene expression in peripheral blood leukocytes with severity of ischemic stroke

International Journal of Neuroscience ◽

10.3109/00207454.2013.837462 ◽

2013 ◽

Vol 124 (4) ◽

pp. 271-280 ◽

Cited By ~ 7

Author(s):

Jia Liu ◽

Jun Yan ◽

Judith M Greer ◽

Stephen J Read ◽

Robert D Henderson ◽

...

Keyword(s):

Gene Expression ◽

Ischemic Stroke ◽

Peripheral Blood ◽

Peripheral Blood Leukocytes ◽

Blood Leukocytes

Download Full-text

LPL methylation of peripheral blood leukocytes - Prophet of atherothrombotic stroke

10.21203/rs.3.rs-16075/v1 ◽

2020 ◽

Author(s):

Minjia Xiao ◽

Zhijie Xiao

Keyword(s):

Ischemic Stroke ◽

Peripheral Blood ◽

Methylation Level ◽

Epigenetic Modification ◽

Hunan Province ◽

Control Group ◽

P Value ◽

Peripheral Blood Leukocytes ◽

Blood Leukocytes ◽

Cpg Dinucleotides

Abstract Background: Lipoprotein lipase (LPL) is a glycoprotein enzyme playing a pivotal role in energy and lipoprotein metabolism. It hydrolyzes triglyceride-rich lipoprotein and produces fatty acids as well as monoacylglycerol. Multiple risk factors including age, sex, hypertension, smoking and hyperlipidemia contribute to ischemic stroke (IS). Hyperlipidemia is an extremely pivotal risk factor for ischemic stroke especially atherothrombotic stroke (ATS) in that it can lead to intracranial or extracranial vessels’ atherosclerosis analogous to atherosclerosis of coronary artery. However, potential epigenetic mechanisms contributing to IS are not been explored thoroughly. The aim of this study was to illuminate relationship among individuals’ peripheral blood leukocytes methylation level of LPL-promoter-CpG dinucleotides, serum lipid and ATS of Chinese Han population in Hunan Province. Methods: Peripheral blood samples were collected from acute atherothrombotic stroke patients and healthy people, and methylation level of cytosine–phosphate–guanine (CpG) island in LPL promoter region was measured by qPCR and pyrosequencing. Biochemical and anthropometric variables have also been taken into consideration.Results: Integral LPL-promoter-CpG dinucleotides methylation level of case group is evidently higher than control group (38.8±8.4% percentile vs 25.7±6.6% percentile, P value=0.000). DNA methylation at the CpG9~16 locus and CpG20~21 locus was related to ATS after adjusting for gender, previous history of diabetes and hypertension, smoking, drinking, body mass index, and blood lipid levels(CpG9 49.3±24.9, 31.3±13.6, P value =0.02; CpG10 61.3±24.5, 34.0±18.4, P value =0.002; CpG11 71.3±17.3, 32.0±21.1, P value =0.000; CpG12 75.3±17.7, 44.7±19.6, P value =0.000; CpG13 72.0±20.4, 50.0±25.4, P value =0.014; CpG14 63.3±18.0, 45.3±22.0, P value =0.021; CpG15 56.7±13.5, 34.7±14.6, P value =0.000; CpG16 50.0±12.5, 31.3±20.0, P value =0.005; CpG20 52.0±13.2, 27.3±20.9, P value =0.001; CpG21 55.3±11.9, 34.0±22.3, P value =0.004). There is no statistically significant connection between either carotid atherosclerosis or gender and methylation level. Besides, we found a negative association between LPL methylation status and HDL-C levels, whereas LPL gene methylation was linked with LDL-C levels and age positively.Conclusion: We illustrate that epigenetic modification of LPL methylation may have an influence on lipids metabolism and occurrence of ATS, which may be a promising indicator for occurrence risk of ischemic stroke. Whether these findings are valid need further warrant and future prospective.

Download Full-text