粘附分子VCAM-1、ICAM-1和PECAM-1单核苷酸多态性与子痫前期的相关性研究

发布时间：2018-04-16 22:18

本文选题：子痫前期 + VCAM-1　；参考：《南方医科大学》2014年硕士论文

【摘要】：研究背景子痫前期是妊娠期特有的疾病,以高血压、蛋白尿、全身内皮细胞激活受损及过度的炎症反应为特征的一组综合征,在我国发病率为9.4%-10.4%。该病病情严重时患者可因抽搐、昏迷、心脑血管意外、心功能衰竭、肺水肿、肝肾功能衰竭、弥漫性血管内出血等而导致死亡；围产儿可出现胎儿生长受限、胎儿宫内窘迫、胎死宫内、早产、新生儿窒息等不良结局,是临床上导致孕产妇及围产儿死亡的重要产科疾病,也是我国孕产妇死亡的第二大原因。半个世纪以来,研究者们尝试从不同角度阐明子痫前期的发病机制,希望通过对该病的病因学研究,找到有效的预测、预防和治疗的方法,然而迄今为止其病因及发病机制仍未清楚,故对于子痫前期的预测、预防仍较为困难。在被广泛认可的各种病因学说中,遗传易感性在子痫前期的发病中可能扮演着重要角色。作为一种复杂性疾病,子痫前期的易感基因可以有很多。目前研究较多的子痫前期的易感基因有：(1)内皮细胞损伤和血管张力改变易感基因；(2)参与脂类代谢的易感基因；(3)遗传血凝基因；(4)人白细胞抗原HLA-DR4基因；(5)线粒体基因等。然而,由于子痫前期的“异质性”特点,基因易感性在不同人种基因组之间、不同地区之间也存在差异性分布,加之基因与基因、基因与环境之间的相互作用,使得寻找易感基因面临很大挑战。粘附分子是介导细胞与细胞之间、细胞与胞外基质间相互结合的细胞表面受体,参与细胞的识别、活化和信号转导,在免疫应答、炎症发生及血栓形成等方面起着重要的作用。血管细胞粘分子-1(vascular cellular adhesion molecule-1VCAM-1)、细胞间粘附分子-1(intercellular adhesion molecule-1, ICAM-1)和血小板内皮细胞粘附分子-1(platelet endothelial cell adhesion molecule-1PECAM-1)是细胞粘附分子中的免疫球蛋白超家族成员。细胞表面粘附分子异常表达可能与子痫前期的发生有重要影响。研究发现在子痫前期患者血中VCAM-1、ICAM-1和PECAM-1的水平较正常对照组有明显差异。其机制可能与VCAM-1、ICAM-1和PECAM-1基因参与的炎症反应导致血管内皮细胞的损伤,进而引起一系列的病理变化有关。VCAM-1、ICAM-1和PECAM-1基因均存在着变异和多态性。这些基因多态性是否会通过影响子痫前期患者血浆水平的表达而影响子痫前期的发生、发展,目前尚不清楚。当前对于子痫前期与粘附分子VCAM-1、ICAM-1及PECAM-1的相关性的研究均局限于蛋白水平,而从基因学角度去探讨其关系的研究,除Kwon和Tabatabai等报道了ICAM-1基因K469E多态性与子痫前期相关性的研究外,尚未见其他文献报道。而且,由于基因多态性的种族差异及地域性差异,结果也可能不同。目的本研究旨在探讨细胞粘附分子VCAM-1基因rs1041163和rs3181092、ICAM-1基因rs5498及PECAM-1基因rs211865545单核苷酸多态性与子痫前期的相关关系,寻找我国南方地区汉族人群子痫前期的易感基因,了解子痫前期的发病机制,为子痫前期的防控和对症性治疗提供依据。方法选择2011年12月至2013年10月南方医院妇产科病房收治的子痫前期患者110例及同一时期健康足月孕妇110例作为对照组。所有入选者均为我国南方地区汉族人群,单胎妊娠且无血缘关系的散发病例,排除辅助生殖技术妊娠、慢性高血压疾病、肾病、胎膜早破、妊娠期糖尿病、糖尿病及抗磷脂抗体阳性等免疫系统性疾病的病史者。收集纳入研究对象的临床指标(年龄、身高、孕前体重、孕产次、入院时的血压、子痫前期的发病孕周或健康足月孕妇的分娩孕周、白细胞计数、血小板计数及24h尿蛋白定量)进行比较分析。采用候选基因法选择VCAM-1基因rs1041163和rs3181092、ICAM-1基因rs5498及PECAM-1基因rs211865545单核苷酸多态性,用聚合酶链反应-限制性片段多态性(PCR-RFLP)方法对这些多态性位点进行基因分型及DNA序列测序法对PCR-RFLP基因分型的准确性进一步验证。运用单因素Logistic回归分析子痫前期的高危因素及多因素Logistic回归分析以校正这些高危因素对基因多态性与子痫前期相关性造成的影响。并且比较子痫前期组中与子痫前期相关联的多态性位点各基因型对其发病孕周、血压、蛋白尿、白细胞及血小板水平的影响。结果 1.通过比较两组研究对象临床资料后发现：孕产妇年龄、孕次、产次及孕前BMI在两组人群中有统计学差异,其中,子痫前期组35岁的高龄孕产妇、初孕妇及初产妇都较对照组明显增多,孕前BMI也较对照组显著增大。 2. rs1041163、rs3181092、rs5498及rs281865545各单核苷酸多态性位点的基因型频率分布都符合Hardy Weinberg平衡定律(p0.05),表明所研究对象来自同一个群体,具有群体代表性。 3. VCAM-1基因rs3181092位点各基因型中,晚发型子痫前期组AA基因型频率(40.3%)和AA+AG基因型频率(91%)高于对照组(22.7%和78.2%),其分布差异在两组均存在显著性(p0.05)；基因型频率相对凤险显示携带AA基因型和AA+AG基因型者发生晚发型子痫前期的风险分别是携带GG基因型者的4.320倍(OR=4.320,95%CI:1.516-12.308)和2.837倍(OR=2.837,95%CI：1.094-7.357)。晚发型子痫前期组A等位基因频率(65.7%)高于对照组(50.5%),其分布差异在两组存在显著性(p0.05)；等位基因频率相对风险显示携带A等位基因者发生晚发型子痫前期的风险是携带G等位基因的1.879倍(OR=1.879,95%CI:1.205-2.928)。 4. PECAM-1基因rs281865545位点各基因型中,早发型子痫前期组CG基因型频率(65.1%)高于对照组(45.5%),其分布差异在两组中均存在显著性(p0.05)；基因型频率相对风险显示携带CG基因型者发生早发型子痫前期的风险是携带CC基因型的2.240倍(OR=2.240,95%CI:1-5.018)。早发型子痫前期组CG+GG基因型频率(74.4%)和G等位基因频率(41.9%)高于对照组(60%和37.3%),但其分布在两组中均无统计学差异(p0.05)。 5. VCAM-1基因rs1041163位点各基因型频率和等位基因频率在子痫前期组和对照组中的分布差异无显著性(p0.05)。ICAM-1基因rs5498位点各基因型中,子痫前期组AG基因型频率(49.1%)高于对照组(41.8%),但其分布在两组中无显著差异(p0.05)；G等位基因频率(27.3%)高于对照组(24.5%),但其分布在两组中亦无统计学差异(p0.05)。 6.通过单因素Logistic回归分析发现,高龄、初孕、初产及孕前BMI是影响子痫前期发生的危险因素,可增加子痫前期发病的风险。经过多因素Logistic回归分析调整高龄、初孕、初产及孕前BMI后,VCAM-1基因rs3181092单核苷酸多态性的AA基因型仍是影响晚发型子痫前期发生的独立危险因素(p0.05)。调整后的基因型频率相对风险显示携带rs3181092单核苷酸多态性AA基因型者发生晚发型子痫前期的风险是携带GG基因型者的3.927倍(OR=3.927,95%CI:1.195-12.905)。经过多因素Logistic回归分析调整高龄、初孕、初产及孕前BMI后,PECAM-1基因rs281865545多态性位点的CG基因型仍然是影响早发型子痫前期发生的独立危险因素(p0.05),调整后的基因型频率相对风险显示携带rs281865545多态性位点的CG基因型者发生早发型子痫前期的风险是携带CC基因型者的4.193倍(OR=4.193,95%CI：1.456-12.079)。 7.白细胞和血小板在子痫前期患者VCAM-1基因rs3181092位点各基因型间的分布差异存在显著性(p0.05),AA基因型携带者的白细胞和血小板水平低于GG基因型和AG基因型携带者,而各基因型间发病孕周、血压及尿蛋白并无明显差异；PECAM-1基因rs281865545位点各基因型间各临床指标差异亦无统计学意义。结论 1.首次发现VCAM-1基因rs3181092单核苷酸多态性AA基因型和AA+AG基因型与我国南方地区汉族人群子痫前期,特别是晚发型子痫前期的发生有关,AA基因型可能是我国南方地区汉族人群晚发型子痫前期的易感基因型,A等位基因的携带者与晚发型子痫前期有较高的遗传易感性。 2.首次发现了PECAM-1基因rs281865545单核苷酸多态性CG基因型与我国南方地区汉族人群子痫前期,尤其是早发型子痫前期的发生有关,CG基因型可能是我国南方地区汉族人群早发型子痫前期的易感基因型。 3. VCAM-1基因rs1041163单核苷酸多态性及ICAM-1基因rs5498单核苷酸多态性与我国南方地区汉族人群子痫前期无关联。 4. VCAM-1基因rs3181092单核苷酸多态性AA基因型可影响子痫前期患者白细胞及血小板的水平。 5.高龄、初孕、初产及孕前BMI也是影响子痫前期发生的危险因素,可增加子痫前期发病的风险；子痫前期是多因素互相影响和共同作用的结果。
[Abstract]:Background of the study

Preeclampsia is a group of syndrome characterized by pregnancy - specific diseases , such as hypertension , proteinuria , activation of systemic endothelial cells and excessive inflammatory response . In China , the incidence rate is 9.4 % - 10.4 % . In severe cases , the patient can cause death due to convulsions , coma , cardiovascular and cerebrovascular accidents , cardiac failure , pulmonary edema , liver and kidney function failure , diffuse vascular hemorrhage , etc .
In half a century , researchers have tried to clarify the pathogenesis of pre - eclampsia and find effective methods of prediction , prevention and treatment .
( 2 ) the susceptible genes involved in lipid metabolism ;
( 3 ) genetic blood coagulation gene ;
( 4 ) HLA - DR4 gene of human leukocyte antigen ;
( 5 ) mitochondrial gene and so on . However , due to the " heterogeneity " in the early - eclampsia , there is a difference distribution between different regions of the gene , and the interaction between the gene and the gene , gene and environment makes it difficult to find the susceptible gene .

Vascular cell adhesion molecule - 1 ( VCAM - 1 ) , intercellular adhesion molecule - 1 ( ICAM - 1 ) and platelet endothelial cell adhesion molecule - 1 ( platelet endothelial cell adhesion molecule - 1 PECAM - 1 ) have a significant effect on the occurrence and development of VCAM - 1 , ICAM - 1 and PECAM - 1 .

Purpose

The purpose of this study was to investigate the relationship between the single nucleotide polymorphism of the VCAM - 1 gene rs1041163 and rs181092 , the ICAM - 1 gene rs5498 and the PECAM - 1 gene , rs2118655 45 , in the early stage of eclampsia .

method

The clinical indexes ( age , height , pre - pregnancy weight , pregnancy birth time , blood pressure during pregnancy , gestational age , gestational age , gestational age , gestational age , gestational diabetes , diabetes mellitus and anti - phospholipid antibody positive ) were analyzed by using single - factor logistic regression analysis .

Results

1 . After comparing the clinical data of the two groups , it was found that the maternal age , the pregnancy time , the birth time and the pre - pregnancy BMI were significantly different in the two groups . Among them , the 35 - year - old pregnant woman , the early pregnant woman and the pregnant woman in the pre - eclampsia group were obviously increased compared with the control group , and the BMI of the pregnant women was also significantly increased compared with the control group .

2 . The genotype frequency distribution of rs1041163 , rs181092 , rs5498 and rs2818655 45 single nucleotide polymorphism loci was in accordance with Hardy ' s equilibrium law ( p0.05 ) , indicating that the subjects were from the same group and were representative of population .

3 . The frequencies of AA genotype ( 40.3 % ) and AA + AG genotype frequencies ( 91 % ) in the early - onset pre - eclampsia group were higher than those in the control group ( 22 . 7 % and 78.2 % ) .
Compared with control group ( OR = 4.320 , 95 % CI : 1.516 - 12.308 ) and 2.837 - fold ( OR = 2.837 , 95 % CI : 1.094 - 7.357 ) , the frequency of late - onset pre - eclampsia ( 65.7 % ) was higher than that of the control group ( 50.5 % ) .
The relative risk of allele frequencies showed that the risk of late onset of onset of late onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of onset of pre - eclampsia was 1.879 times ( OR = 1.879 , 95 % CI : 1.205 - 2.928 ) carrying the G allele .

4 . The frequency of CG genotype in early - onset pre - eclampsia group ( 65.1 % ) was higher than that in the control group ( 45.3 % ) , and the distribution difference was significant ( p < 0.05 ) .
The relative risk of genotype frequency showed that the risk of early onset of pre - eclampsia was 2.240 times ( OR = 2.240 , 95 % CI : 1 - 5.018 ) of CC genotype . The frequency of CG + GG genotype ( 74.4 % ) and G allele frequency ( 41.9 % ) in early - onset pre - eclampsia group were higher than those in control group ( 60 % and 37.3 % ) , but their distribution was not statistically significant in both groups ( p . 05 ) .

5 . There was no significant difference in the frequency and allele frequencies of the rs1041163 locus of VCAM - 1 gene in the pre - eclampsia group and the control group ( p < 0.05 ) . The frequency of the AG genotype in the early - eclampsia group ( 49.1 % ) was higher than that in the control group ( 41.8 % ) , but the distribution of the distribution of the genotype frequencies and allele frequencies in the early - eclampsia group was higher than that in the control group ( 41.8 % ) .
The frequency of G allele ( 27.3 % ) was higher than that of the control group ( 24.5 % ) , but its distribution was not statistically significant in both groups ( p < 0.05 ) .

6 . A single - factor logistic regression analysis showed that the risk factors of early onset of pre - eclampsia were the age , early pregnancy , early pregnancy and pre - pregnancy BMI , which could increase the risk of pre - eclampsia . After multivariate logistic regression analysis , the genotype of the single nucleotide polymorphism of rs2818655 was 3.927 times ( OR = 3.927 , 95 % CI : 1.195 - 12.905 ) . The risk of genotype frequency after adjustment showed that the risk of early - onset pre - eclampsia was 4.193 times that of CC genotype ( OR = 4.193 , 95 % CI : 1.456 - 12.079 ) .

7 . There was a significant difference in the distribution of VCAM - 1 gene rs181092 in patients with pre - eclampsia ( p < 0.05 ) . The levels of leukocyte and platelet in AA genotype carriers were lower than those of GG genotype and AG genotype carriers , but there was no significant difference in the gestational weeks , blood pressure and urinary protein among the genotypes .
There was no significant difference in clinical parameters between the genotypes of PECAM - 1 and rs2818655 .

Conclusion

1 . The single nucleotide polymorphism AA genotype and AA + AG genotype of the VCAM - 1 gene were found to be related to the pre - eclampsia of Han population in the southern region of China , especially in the pre - eclampsia , AA genotype could be the susceptible genotype of late - onset pre - eclampsia in the Han population in the southern region of China .

2 . The polymorphism of the single nucleotide polymorphism of the PECAM - 1 gene rs2818655 45 was first found to be related to the pre - eclampsia of Han population in the southern region of China , especially in the early - onset pre - eclampsia , and the CG genotype could be the susceptible genotype of early - onset pre - eclampsia in the Han population in the southern region of China .

3 . The single nucleotide polymorphism of the VCAM - 1 gene rs1041163 and the single nucleotide polymorphism of the ICAM - 1 gene rs5498 were not associated with the pre - eclampsia in the Han population in the southern region of China .

4 . The single nucleotide polymorphism of the single nucleotide polymorphism of the VCAM - 1 gene rs181092 can affect the level of white blood cells and platelets in patients with pre - eclampsia .

5 . Age , early pregnancy , early birth and pre - pregnancy BMI are also the risk factors which affect the pre - eclampsia , which can increase the risk of pre - eclampsia .
Preeclampsia is the result of multi - factor interaction and coaction .

【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.244

【参考文献】