Study On The Relationship Between Single Nucleotide Polymorphisms In The Promoter Region Of ITLN-1 And Acute Ischemic Stroke

Objective: Omentin is a kind of adipose tissue-derived cytokine encoded by ITLN1.Researches have indicated that the expression level of circulating omentin-1 is closely related to acute ischemic stroke and its risk factors including diabetes,atherosclerosis,and vasculitis and so on.However,it is remain unclear about the changes of omentin levels in acute ischemic stroke.Now we will implement a case-control study to explore the changes of serum omentin-1 level in patients with acute ischemic stroke compared with non-acute ischemic stroke patients,and to explore whether there are single nucleotide polymorphism(SNP)sites in the promoter region of ITLN-1 that are associated with the risk of acute ischemic stroke.So that can provide a research target for the prevention and treatment of acute ischemic stroke.Methods: First,we collected 90 samples(Case: 44 patients with acute ischemic stroke;Control: 46 non-stroke patients)from the Department of Neurology of Xi’an third Hospital and Xi’an first Hospital during July 2019 to October 2020.The demographic data and medical records of the patients were collected,and the peripheral venous blood of the patients was extracted as biological samples within 24 hours of admission.Enzyme-linked immunosorbent assay(ELISA)was performed to measure the expression levels of serum omentin-1 both in the case group and the control group to explore the changes of serum omentin-1 levels in patients with acute ischemic stroke.Furthermore,the genomic DNA was extracted from peripheral blood leukocytes,and the full-length DNA fragment in the promoter region of ITLN-1 was amplified by polymerase chain reaction(PCR).After the amplified products were identified by agarose gel electrophoresis,we carried out Sanger sequencing method to identify these sequence of products.The sequence was compared with the reference sequence of the promoter region of ITLN-1 to determine the SNP locus.The t-test and chi-square test were used to analyze the changes of serum omentinin-1 level in patients with acute cerebral infarction.Fisher’s exact test and logistic regression model were used to analyze the correlation between single nucleotide polymorphisms in the promoter region of omentinin gene and various disease phenotypes.Results: 1.In Xi’an,Shaanxi Province,the number of smokers and the expression level of blood HCY in the acute ischemic stroke group are higher than those in the control group,and the expression level of serum omentin-1 in the acute ischemic stroke group is lower compared with the control group,these differences were statistically significant(P < 0.05).2.A total of 19 common single nucleotide polymorphisms were detected in the DNA sequence of the promoter region of ITLN-1 in Xi’an,Shaanxi Province.Including: 1 rs60721814(G >C),2 rs80262455(C > T),3 rs58615002(A > G),4 rs111321091(C > T),5 rs58170012(A > G),6 rs4656958(C > T),7 rs74547401(A > G),8 rs7411035(A > C),9 rs12091185(G > A),10 rs12094703(T > C),11 rs6427553(A > G),12 rs12094649(T > C),13rs12091040(T > A),14.17-268(G > A),18 rs79209815(G > A),19 rs77824633(G > A).3.Independent of hypertension,atrial fibrillation,diabetes,coronary heart disease,blood total cholesterol level,blood triglyceride level,blood high density lipoprotein level,blood low density lipoprotein level and blood HCY level and other confounding factors,the mutation of rs12091185(G >A)tends to decrease the risk of acute ischemic stroke.The mutation of rs60721814(G >C)、 rs80262455(C >T)、 rs58615002(A >G)、 rs111321091(C >T)、rs58170012(A >G)、 rs12094703(T >C)、 rs12094649(T >C)、 rs12091040(T >A)、rs60429964(G >C)、rs77824633(G >A)tends to increase the individual’s risk of acute ischemic stroke.Conclusion: In the population of Xi’an,Shaanxi Province,smoking,higher blood homocysteine levels,and lower serum omentin-1 levels were associated with a higher risk of acute ischemic stroke.There are 19 common single nucleotide polymorphisms in the promoter region of ITLN-1,the risk of acute ischemic stroke tends to be decreased with the mutation of the rs77824633(G >A),while the risk tends to be increased with the mutation of rs60721814(G >C)、rs80262455(C >T)、rs58615002(A >G)、rs111321091(C >T)、rs58170012(A >G)、rs12094703(T >C)、rs12094649(T >C)、rs12091040(T >A)、rs60429964(G >C)and rs77824633(G >A).