eNOS、EPO、TNF-α基因多态性与青海省汉族妊娠高血压疾病相关性研究

发布时间：2018-04-24 11:05

本文选题：妊娠高血压疾病 + 单核苷酸多态性　；参考：《青海大学》2017年硕士论文

【摘要】：目的通过研究内皮型一氧化氮合酶(endothelial nitricoxide synthase,e NOS)基因G894T、促红细胞生成素(erythropoietin,EPO)基因T3541G、肿瘤坏死因子-α(tumour necrosises factor-alpha,TNF-α)基因G308A等多态性位点基因型频率在青海省汉族妊娠高血压疾病(pregnancy-induced hypertension syndrom,PIH)患者和正常孕妇之间分布的差异性,检测两组血浆中EPO、TNF-α含量,比较两组血液中e NOS m RNA相对表达差异性,探讨e NOS基因G894T、EPO基因T3541G、TNF-α基因G308A多态性与妊娠高血压疾病的关联性,为青海地区妊娠高血压疾病的预防提供依据。方法收集青海省各大医院汉族人群中妊娠高血压患者和正常孕妇外周血标本和临床资料,提取基因组DNA,采用限制性片段长度多态性聚合酶链反应(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)方法分别扩增e NOS SNP/G894T、EPO SNP/T3541G、TNF-αSNP/G308A等多态性位点,采用特异性酶对e NOS基因G894T、EPO基因T3541G、TNF-α基因G308A进行多态性分型。分别对不同基因型的目的片段测序,将测序结果在美国国立生物技术信息中心(national center for biotechnology information,NCBI)数据库中比对,确保扩增产物真实可靠,并对酶切结果加以验证;使用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测血浆中EPO、TNF-α含量;使用半定量反转录-聚合酶链反应(semi-quantitative reverse transcription and polymerase Chain reaction,Sq RT-PCR)方法分析血液中e NOS基因m RNA表达情况。结果1.e NOS基因GG、GT、TT基因型频率在PIH组分别为17.4%、82.6%、0%,对照组分别为0%、95.6%、4.4%,两组比较存在差异性(P0.05)。PIH组G、T等位基因频率分别为58.7%、41.3%,对照组分别为47.8%、52.2%,两组比较差异有统计学意义(P0.05),PIH组G等位基因频率高于对照组。PIH组比对照组(OR=1.553,95%CI为1.107~2.178),G等位基因为PIH的易感基因(OR=1.229,95%CI为1.048~1.441)。2.EPO基因TT、TG、GG基因型频率在PIH组分别为9.7%、74.2%、6.1%,对照组分别为42.9%、46.4%、10.7%,两组比较有差异(P0.05)。PIH组野生纯合型(TT)频率明显低于对照组,突变型(TG、GG)基因型频率高于对照组。PIH组等位基因T、G频率分别为46.8.%,53.2%,对照组分别为66.1%、33.9%,两组比较差异有统计学意义(P0.05),PIH组T等位基因频率低于对照组,G基因频率明显高于对照组,PIH比对照组(OR=0.451,95%CI为0.311~0.655),G等位基因为妊高征的危险因素(OR=1.569,95%CI为1.263~1.949)。3.TNF-α基因GG、GA、AA基因型频率在PIH组中分别为18.18%、72.73%、9.09%,对照组为42.86%、52.38%、4.76%,两组比较有显著差异性(P0.01)。PIH组突变型(GA+AA)基因型频率(81.82%)明显高于对照组(57.14%)。PIH组G等位基因频率(54.55%)低于对照组(69.05%),A等位基因频率(45.45%)高于对照组(33.95%),(P0.01)。PIH组比对照组(OR=0.538,95%CI:0.375~0.771),A等位基因为PIH的易感基因(OR=1.469,95%CI:1.170~1.843)4.PIH组血浆中EPO含量为(454.113±35.097)pg/m L,对照组为(396.316±50.262)pg/m L,PIH组明显高于对照组(P0.01)。5.TNF-α含量在PIH组血浆中为(21.976±17.570)pg/m L,对照组为(27.797±14.730)pg/m L两组比较没有统计学差异(P0.05)。6.PIH组e NOS m RNA表达相对强度为(1.007±0.155),对照组为(1.243±0.329),PIH组低于对照组(P0.05)。结论1.e NOS SNP/G894T、EPO SNP/T3541G、TNF-αSNP/G308A多态性与青海省汉族妊娠高血压疾病相关,e NOS SNP/G894T位点G等位基因、EPO SNP/T3541G位点G等位基因、TNF-αSNP/G308A位点A等位基因为妊娠高血压疾病的易感基因。2.EPO基因在妊娠高血压疾病患者血浆中高表达。3.TNF-α含量在PIH患者血浆中没有变化。4.e NOS m RNA在PIH患者血液中低表达。
[Abstract]:Objective to study the genotype frequencies of the polymorphic loci, such as endothelial nitricoxide synthase (E NOS) gene G894T, erythropoietin (erythropoietin, EPO) gene, and tumor necrosis factor - alpha (tumour necrosises factor-alpha, alpha) gene polymorphic loci. The difference in distribution between nancy-induced Hypertension Syndrom, PIH) and normal pregnant women, the content of EPO, TNF- alpha in two groups of plasma, and the relative expression of E NOS m RNA in the two groups, and the association between the e NOS gene, the polymorphism of the alpha gene and the pregnancy induced hypertension, and the pregnancy induced pregnancy in Qinghai region Methods to provide basis for the prevention of pregnancy induced hypertension. Methods to collect the peripheral blood samples and clinical data of pregnant hypertension and normal pregnant women from the Han population of Qinghai Province, and to extract genomic DNA and use the restrictive fragment length polymorphism polymerase chain reaction (polymerase chain reaction-restriction fragment length polymorphism, PCR). -RFLP) the polymorphic loci, such as e NOS SNP/G894T, EPO SNP/T3541G, TNF- alpha SNP/G308A, were amplified by the method, and the polymorphic typing was carried out by specific enzymes to e NOS gene G894T, EPO gene and alpha gene. For biotechnology information, NCBI) comparison in the database to ensure the authenticity of the products, and to verify the results of the enzyme digestion; using the enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA) to detect the content of EPO, TNF- a in plasma, and the use of Semidefinite reverse transcription polymerase chain reaction (Semi-quantitative reverse) Iption and polymerase Chain reaction, Sq RT-PCR) method analysis of E NOS gene m RNA expression in blood. Results 1.e genes were 17.4%, 82.6%, 0%, respectively 0%, 95.6%, 4.4% in the control group, and two groups were 58.7%, 41.3%, and control components, respectively. There were significant differences in the 47.8%, 52.2%, and two groups (P0.05). The frequency of G alleles in group PIH was higher than that in the control group (OR=1.553,95%CI was 1.107~2.178), and the G allele was PIH's susceptibility gene (OR=1.229,95%CI 1.048~1.441).2.EPO gene TT, and the frequency of genotype was 9.7%, 74.2%, 6.1%, and control component, respectively. The frequency of wild homozygous type (TT) in group.PIH was significantly lower than that of control group in group 42.9%, 46.4%, 10.7% and two. The frequency of mutant type (TG, GG) genotype was higher than that of.PIH group T in the control group, the frequency of G was 46.8.%, 53.2%, and the control group was 66.1%, 33.9% and two, respectively (P0.05), PIH group T allele frequency. The frequency of G gene was significantly higher than that in the control group, PIH was higher than the control group (OR=0.451,95%CI 0.311~0.655), G allele was the risk factor of pregnancy induced hypertension (OR=1.569,95%CI 1.263~1.949).3.TNF- a gene GG, GA, AA genotype frequencies were 18.18%, 72.73%, 9.09% respectively in the PIH group, 42.86%, 52.38%, 4.76% in the control group, and two groups were compared with the two groups. The genotype frequency of P0.01.PIH group (81.82%) was significantly higher than that of control group (57.14%).PIH group G allele frequency (54.55%) was lower than that of control group (69.05%), A allele frequency (45.45%) was higher than that of control group (33.95%), (P0.01).PIH group than control group (OR=0.538,95%CI:0.375~0.771), A allele was PIH susceptible gene (OR=1.4) 69,95%CI:1.170~1.843) the content of EPO in plasma of 4.PIH group was (454.113 + 35.097) pg/m L, the control group was (396.316 + 50.262) pg/m L, PIH group was significantly higher than that of control group (P0.01).5.TNF- alpha in PIH group plasma (21.976 + 17.570) pg/m, and the control group was (27.797 + 14.730). The relative intensity of the expression was (1.007 + 0.155), the control group was (1.243 + 0.329), and the PIH group was lower than the control group (P0.05). Conclusion 1.e NOS SNP/G894T, EPO SNP/T3541G, TNF- alpha SNP/G308A polymorphism is associated with pregnancy induced hypertension in Qinghai Han nationality, e NOS SNP/G894T locus allele. The high expression of.2.EPO gene in the plasma of hypertensive disorder complicating pregnancy with high expression of.3.TNF- alpha in the plasma of patients with pregnancy induced hypertension does not change the low expression of.4.e NOS m RNA in the blood of PIH patients.

【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.246

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