| Objective:Acute ischemic stroke(AIS),a kind of cerebrovascular diseases caused by environmental and hereditary factors,is characterized by high morbidity,mortality,disability and recurrence,and puts a heavy burden on society and family.However,the specific pathogenesis of AIS is unclear till now.In recent years,studies have shown that microRNAs(miRNAs)and their single nucleotide polymorphisms may be involved in the pathophysiological processes of the occurrence,development and prognosis of AIS,among which miRNA-146a is one of the most concerned miRNAs.The purpose of this study was to investigate the expression of miRNA-146a in human peripheral blood mononuclear cells in patients with AIS and the association between single nucleotide polymorphisms and genetic susceptibility of AIS,so as to provide molecular biological evidence for the prevention and treatment of AIS.Methods:A total of 137 inpatients diagnosed with AIS in the Department of Neurology,Affiliated Hospital of North Sichuan Medical College from July 2020 to December 2020 were selected as the experimental group,and 100 healthy subjects were selected as the control group during the same period.Patients in the experimental group were divided into mild Stroke group(NIHSS≤3 points)and non-mild Stroke group(NIHSS>3 points)according to the National Institutes of Health Stroke Scale(NIHSS)score at admission.According to the Trial of Org10172 in Acute Stroke Treatment(TOAST)classification,the AIS was divided into large artery atherosclerosis(LAA),small vessel occlusion(SAA),cardioembolism(CE),stroke of other etiology(SOE)and stroke of unknown etiology(SUE).According to the course of disease at admission,the patients were divided into:course ≤1 day(T1)group,1 day<course ≤3 days(T2)group,3<course<7 days(T3)group,7<course ≤14 days(T4)group.The peripheral venous blood and relevant clinical data of the experiment and control group were collected.Peripheral blood mononuclear cells(PBMCs)were Isolated and extracted.Real-time quantitative Reverse Transcription Polymerase Chain Reaction(RT-qPCR)was used to detect the relative expression of miRNA-146a in PBMCs.Meanwhile,the whole genome DNA was extracted by DNA extraction kit,and the distribution of different alleles and genotypes of miRNA-146a rs2910164 in the experimental and control groups was detected by TaqMan-MGB real-time fluorescence quantitative PCR.SPSS 23.0 software was used to analyze the experimental data.Results:(1)Comparison of basic data between the experimental group and the control group:there was no statistical significance in age and gender between the experimental group and the control group(P>0.05).The incidence of drinking history,smoking history,hypertension and diabetes in the experimental group were higher than those in the control group(P<0.05).The percentage of leukocytes and neutrophils in the experimental group is higher than that in the control group;The percentages of leukocytes and neutrophils in the experimental group were significantly higher than those in the control group(P<0.001),and the percentages of lymphocytes,monocytes,hemoglobin and erythrocytes in the experimental group were significantly lower than those in the control group(P<0.05).The incidence of atrial fibrillation,carotid artery plaque,pulmonary infection,deep vein thrombosis,urinary tract infection and post-stroke depression were all significantly higher in the non-mild stroke group than in the mild stroke group(P<0.05).Meanwhile,the white blood cell and high sensitivity C-reactive protein levels in non-mild stroke group were significantly higher than those in mild stroke group(P<0.05);the percentage of lymphocyte and hemoglobin in non-mild stroke group were lower than those in mild stroke group(P<0.05).In the TOAST classification,the average age of SOE type cerebral AIS was significantly lower than that of other groups(P=0.00).The incidence of hypertension and diabetes in LAA and SAA types was higher than that in CE and SOE types;the incidence of carotid plaque in LAA and CE types was significantly higher than that in SOE type;the incidence of atrial fibrillation in CE type was significantly higher than that in other types(P<0.05).The incidence of pulmonary infection and post-stroke depression in CE type was higher than that in SAA type,and the incidence of urinary tract infection in LAA type was higher than that in SAA type,and the differences were all statistically significant(P<0.05).The incidence of non-mild stroke was higher in LAA and CE types than SAA type(P<0.05).There was no significant statistical difference in the TOAST classification as for smoking history,drinking history and stroke history(P>0.05).For the course groups,there was no significant difference in age,gender,smoking history,drinking history,hypertension,diabetes and carotid plaque(P>0.05).(2)Comparison of miRNA-146a expression levels in the experimental group and the control group:the relative expression level of miRNA-146a in the experimental group(1.65±0.11)was significantly higher than that in the control group(1.13±0.09,P=0.002).In the subgroup analysis,The relative expressions of miRNA-146a in different course of IS are T1(1.26±0.19),T2(1.14±0.11),T3(2.04 ±0.26)and T4(2.07±0.23),respectively.The level of miRNA-146a in T3 and T4 groups was significantly higher than that in T1,T2 groups and the control group(P<0.05),while there was no statistical significance between T1 and T2 groups and the control group about the relative expression level of miRNA-146a(P>0.05),and there was no significant difference between T1 and T2 groups,T3 and T4 groups(P>0.05).In addition,there was no significant difference in the relative expression of miRNA-146a between mild stroke group(1.69±0.15)and non-mild stroke group(1.61±0.17)(P>0.05),and there was no significant difference in the relative expression of miRNA-146a between different TOAST subtypes(P>0.05).(3)There was no statistical difference of rs2910164C/G polymorphism loci between the experiment and control groups(P=0.703,OR=0.930,95%CI=0.641-1.349)for distribution of miRNA-146a rs2910164C/G polymorphism.There was no association between rs2910164C/G polymorphism and AIS genetic susceptibility after adjusting for age and sex(P>0.05).However,among the TOAST subtypes,the frequency of GG genotypes in patients with SOE type AIS was higher than in the control group(P=0.00,OR:4.825,95%CI:2.720-8.562).At the same time,miRNA-146A rs2910164C/G polymorphism was associated with genetic susceptibility to ischemic stroke in the heterozygous and homozygous comparison model,according to stratified analysis of the severity of stroke at admission(P=0.024,OR=3.163,95%,CI=1.163-8.607;P=0.011,OR=0.266,95%,CI=0.095-0.739).We failed to find the relative expression of miRNA-146a was influenced by different genotypes of miRNA-146a rs2910164 polymorphism locus(P>0.05).Conclusions:(1)The incidence of non-mild AIS was higher in patients with atrial fibrillation and carotid plaque,and non-mild stroke is more prone to pulmonary infection,deep vein thrombosis,urinary tract infection,post-stroke depression and other complications.Patients with SOE type AIS were younger than those with other types of AIS.Patients with hypertension and diabetes were prone to LAA type and SAA type AIS.Patients with carotid artery plaques were prone to LAA type and CE type AIS.LAA type and CE type AIS were more prone to pulmonary infection,post-stroke depression,urinary tract infection and other complications than SAA type AIS.In addition,LAA and CE types were more likely to be manifestated non-mild AIS than SAA types.These findings provided reference for the assessment of AIS severity and the prevention and treatment of its complications.These findings provide a reference for the assessment of the severity of AIS and the prevention and treatment of its complications.The percentages of white blood cells and neutrophils were increased in patients with AIS,and the levels of super-sensitive C-reactive protein and white blood cells were significantly increased in non-mild AIS,suggesting that AIS may be related to acute inflammatory response,and the above indicators were expected to be used as laboratory indicators for the diagnosis and the severity evaluation of of AIS.(2)The expression level of miRNA-146a was significantly increased in patients with AIS,and there were differences among patients with different disease courses.The expression level of miRNA-146a was significantly increased in patients with disease course 3-14 days compared with patients with disease course<3 days,suggesting that miRNA-146a may be involved in the pathophysiological process of AIS occurrence and development,and was expected to become one of the treatment targets of AIS.(3)MiRNA-146a rs2910164C/G polymorphism may not be associated with genetic susceptibility to AIS,but rs2910164C/G polymorphism may be associated with the severity of AIS at admission,and rs2910164 GG genotype may increase the risk of SOE type stroke.In the future,multi-center,large-sample and prospective studies are needed to further clarify the correlation. |
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