维生素D代谢通路相关基因多态与孕早期血浆维生素D水平的关联研究

发布时间：2018-04-28 20:48

本文选题：孕妇 + 血浆25(OH)D　；参考：《浙江大学》2017年硕士论文

【摘要】：研究背景及目的维生素D缺乏是一个全球性的健康问题,全世界有超过十亿人维生素D缺乏或者不足。维生素D缺乏与多种骨骼肌肉相关疾病和非骨骼肌肉疾病发生相关。由于女性户外活动较少且常懫取防晒措施,而且孕妇在孕期维生素D的需求明显增加,所以维生素D缺乏在妊娠期孕妇中非常普遍。孕妇在妊娠期维生素D的缺乏或不足可能与先兆子痫、妊娠期糖尿病、炎症性疾病等妊娠期并发症和早产、低出生体重等不良妊娠结局相关,甚至还会影响母亲和子代远期疾病的发生,包括癌症、糖尿病、炎症疾病、过敏性疾病、神经退行性疾病等。血浆25(OH)D被认为是代表体内维生素D营养状况的最佳指标。环境因素包括季节、地理纬度、维生素D摄入量等,以及遗传因素都会影响体内的维生素D水平。家庭研究和双生子研究发现血浆25(OH)D水平变异中遗传因素估计约占23-80%。既往研究发现NADSYN1、DHCR7、GC、CYP2R1和VDHR等维生素D代谢通路相关基因的单核苷酸多态性与血浆25(OH)D水平存在显著的关联。但是研究对象主要是普通成年人,且结论不一致。目前,在孕妇人群中尚未见对维生素D代谢通路相关基因的单核苷酸多态性与孕期血浆25(OH)D水平之间的关联进行的系统性的研究。因此,本研究以浙江省舟山市妇幼保健院为研究现场,采用横断面研究的设计探讨孕妇维生素D代谢通路相关基因(NADSYN1、DHCR7、GC、CYP2R1、CYP3A4、CYP27A1、CYP27B1、CYP244A1、VDHR)多态性与孕早期血浆 25(OH)D 水平之间的关联;并且对可能的基因-环境及基因-基因的交互作用与血浆25(OH)D水平之间的关联进行分析,为孕期合理补充维生素D提供更多的科学依据。材料和方法本研究以浙江省舟山市妇幼保健院为研究现场,在知情同意后,通过面对面访谈获得流行病学调查问卷资料(包括研究对象的一般社会人口学特征、生活行为方式、身高体重等基本信息),并采集孕早、中、晚期空腹血样。从2011年8月到2014年4月期间纳入队列的孕妇中,随机选择759例已完成分娩的孕妇,检测其孕早期血浆25(OH)D水平,并且利用孕早期白细胞血样检测相关基因多态。本研究中,懫用高效液相色谱串联质谱分析法测定孕早期血浆25(OH)D水平。通过文献阅读、数据库检索等方法选择基因的SNP位点并且运用基于MALDI-TOF-MS技术的美国Sequenom公司的MassARRAY系统完成SNP分型。采用多元线性模型分析、叉生分析、多因子降维分析等方法,对基因多态的主效应、基因-环境、基因-基因交互作用与血浆25(OH)D水平之间的关联进行分析。结果本研究纳入的759名孕早期孕妇的平均年龄为28.30±3.06岁,平均孕周为11.51±0.63周,在春季、夏季、秋季和冬季采血人数分别为168、192、195和204人,分别占总人数的22.13%、25.30%、25.69%和26.88%。孕妇的血浆25(OH)D水平平均为 15.64 ± 6.52 ng/mL,其中 593 人维生素 D 缺乏(25(OH)D20 ng/mL),另外 166 人的血浆 25(OH)D ≥ 20 ng/mL,.分别占 78.13%和 21.87%。采用线性回归模型分析基因多态与血浆25(OH)D水平的关联,在检测的.49个SNPs中,其中10个SNP与血浆25(OH)D水平显著相关。在GC基因的多态分析中,经年龄、孕前BMI、采血季节、体育锻炼情况调整后,与rs16846876的AA基因型相比,AT基因型和TT基因型携带者的血浆25(OH)D水平分别平均降低1.22 ng/mL 和 2.72ng/mL(P=0.0121;P=0.0006);与 rs17467825 的 AA 基因型携带者相比,GA基因型携带者和GG基因型携带者的血浆25(OH)D水平也降低(β =-1.95,P0.0001;β=-2.88,P= 0.0003)。结果还显示,与 rs17467825 连锁强度很强(r20.98)的 SNPs 包括 rs2282679、rs3755967、rs2298850、rs4588 的多态均与血浆25(OH)D水平相关,并且其基因型与血浆25(OH)D水平的相关趋势与rs17467825的相似。另外,与rs7041的AA基因型相比,CA基因型携带者和CC基因型携带者的血浆25(OH)D水平平均升高1.80 ng/mL和2.30 ng/mL(P =0.0002;P=0.0123);与rs1155563的TT基因型相比,CC基因型的携带者血浆25(OH)D水平较低(ββ=-1.80,P=0.0104);而rs2298849的GG基因型携带者与AA基因型携带者相比,血浆25(OH)D水平相对较高(β=1.44,P=0.0423)。另外在CYP3A4基因多态分析中,与rs2242480的CC基因型相比,TT基因型的携带者血浆25(OH)D水平较高(ββ=2.48,P=0.0081)。单体型分析结果显示,与GC基因单体型TTGTGTCA相比,调整可能的混杂因素后,单倍型 ATAGGCTG(β=2.17,P =0.0041)、ATATGCTG(β=1.47,P =0.0110)、ACATTCTG(β=3.08,P=0.0001)、ATATTTCA(β=1.99,P=0.0092)均显著增加血浆25(OH)D水平。LP2基因中,经调整后,与最常见单倍型TCCCGCA相比,TCCCCTG单倍型显著降低血浆25(OH)D水平(β=-1.55,P=0.0462)。但未发现其他基因单倍型分析中与血浆25(OH)D水平存在显著关联。基因-环境、基因-基因交互作用的叉生分析结果显示,采血季节与rs933994-CYP27A1、rs2246709-CYP3A4多态性的交互作用1与血浆25(OH)D水平存在关联。此外,本研究还发现 rs7041-GC 与 rs1562902-CYP2R1、rs10766197-CYP2R1、rs2248137-CYP24A1、rs2242480-CYP34;rs2298849-GC与 rs2246709-CYP3A4;rs2209314-CYP24A1与rs1544410-VDR之间的交互作用与血浆25(OH)D水平存在显著的关联。GMDR分析基因多态交互效应发现,由rs7041-GC、rs1155563-GC,rs1562902-CYP2R1,rs2209314-CYP2441构成的模型是预测血浆25(OH)D水平的最佳模型。结论在孕妇人群中,维生素D代谢通路基因多态性与孕早期血浆25(OH)D水平存在关联,其中GC基因变异可能是影响孕早期血浆25(OH)D水平的主要基因变异。采血季节与维生素D代谢通路基因对维生素D水平存在交互作用。研究结果仍需要进一步予以验证。
[Abstract]:Background and objective vitamin D deficiency is a global health problem, more than one billion people worldwide are deficient in or deficient in vitamin D. Vitamin D deficiency is associated with a variety of skeletal muscle related diseases and non skeletal muscle diseases. The demand for vitamin D deficiency is very common in pregnant women. The deficiency or deficiency of vitamin D during pregnancy may be associated with pregnancy complications such as preeclampsia, gestational diabetes, inflammatory disease, premature birth, low birth weight, and even the mother and progeny disease. The occurrence of cancer, diabetes, inflammation, anaphylaxis, neurodegenerative diseases, and so on. Plasma 25 (OH) D is considered to represent the best indicator of vitamin D nutrition in the body. Environmental factors including season, geographical latitude, vitamin D intake, and genetic factors affect the level of vitamin D in the body. Family studies and double The study found that genetic factors in plasma 25 (OH) D levels were estimated to account for about 23-80%. previously found in NADSYN1, DHCR7, GC, CYP2R1 and VDHR, and there was a significant correlation between the single nucleotide polymorphisms of the vitamin D metabolic pathway related genes and plasma 25 (OH) D levels. However, the research subjects were mainly common adults, and the conclusions were inconsistent. Currently, There is no systematic study of the association between single nucleotide polymorphisms of vitamin D pathway related genes and plasma 25 (OH) D levels in pregnant women. Therefore, this study took the maternal and child health care hospital in Zhoushan, Zhejiang as a study site, and used a cross-sectional study to explore the correlation of vitamin D metabolic pathways in pregnant women The association between gene polymorphism (NADSYN1, DHCR7, GC, CYP2R1, CYP3A4, CYP27A1, CYP27B1, CYP244A1, VDHR) and plasma 25 (OH) D levels in the early pregnancy; and the association between the interaction of the possible gene environment and gene gene and the level of plasma 25 (OH), providing more scientific evidence for the rational supplement of vitamin supplements during pregnancy. Materials and methods were studied on the site of maternal and child health care hospital in Zhoushan, Zhejiang. After informed consent, through face-to-face interviews, the epidemiological survey data (including the general social demography characteristics, lifestyle, height and weight and other basic information) were obtained, and the early, middle and late fasting blood samples were collected from 2011. During the period from August to April 2014, 759 pregnant women who had been given birth were randomly selected to test the level of plasma 25 (OH) D at the early stage of pregnancy and the related gene polymorphisms in the early pregnancy. In this study, the level of plasma 25 (OH) D in early pregnancy was measured by HPLC MS analysis. Reading, database retrieval and other methods selected gene SNP loci and using the MassARRAY system of American Sequenom company based on MALDI-TOF-MS technology to complete SNP typing. Multiple linear model analysis, cross analysis, multi factor reduction analysis and other methods, the main effect of gene polymorphism, gene environment, gene gene interaction and plasma 25 ( OH) the correlation between D levels was analyzed. Results the average age of 759 pregnant women in this study was 28.30 + 3.06 years old, and the average gestational weeks were 11.51 + 0.63 weeks. The number of blood samples in spring, summer, autumn and winter were 168192195 and 204, respectively, which accounted for 22.13%, 25.30%, 25.69% and 26.88%. in the total number of pregnant women, respectively, and the levels of plasma 25 (OH) D in pregnant women. The mean was 15.64 + 6.52 ng/mL, of which 593 were vitamin D deficiency (25 (OH) D20 ng/mL), and 25 (OH) D > 20 ng/mL in the other 166, respectively, and 78.13% and 21.87%., respectively, using linear regression model to analyze the association between gene polymorphism and plasma 25 (OH) D level, and 10 of them and plasma 25. In the GC gene polymorphism analysis, the plasma 25 (OH) D levels of the AT genotypes and TT genotype carriers decreased by 1.22 ng/mL and 2.72ng/mL (P=0.0121; P=0.0006), compared with the AA genotype of rs16846876, after age, pre pregnancy, blood harvest season, and physical exercise, and compared with the AA genotype of the AT genotype and TT genotype carriers. The level of plasma 25 (OH) D of the carriers of the type carrier and the GG genotype also decreased (beta =-1.95, P0.0001; beta =-2.88, P= 0.0003). The results also showed that SNPs including rs2282679, rs3755967, and P= 0.0003 was strongly associated with the level of blood plasma 25. In addition, the level of plasma 25 (OH) D in CA genotype carriers and CC genotype carriers increased by 1.80 ng/mL and 2.30 ng/mL (P =0.0002, P=0.0123), compared with the AA genotype of rs7041, and the level of plasma carriers in the carriers of the genotype was lower than that of the AA genotype (Beta beta); In rs2298849 GG genotype carriers, plasma 25 (OH) D levels were relatively higher than those of AA genotype carriers (beta =1.44, P=0.0423). In CYP3A4 gene polymorphism analysis, the level of carrier plasma 25 (OH) was higher than the CC genotype of rs2242480 (beta beta). After adjusting the possible confounding factors, the haplotype ATAGGCTG (beta =2.17, P =0.0041), ATATGCTG (beta =1.47, P =0.0110), ACATTCTG (beta =3.08, P=0.0001) all significantly increased the plasma 25 (beta =3.08, P=0.0001), and, after adjustment, compared with the most common haplotype, the haplotype significantly reduced the plasma level of 2. 5 (OH) D level (beta =-1.55, P=0.0462). But there was no significant association with plasma level 25 (OH) D in other gene haplotypes. Gene environment, gene gene interaction, cross analysis showed that the interaction between the blood harvest season and rs933994-CYP27A1, rs2246709-CYP3A4 polymorphisms was associated with plasma 25 (OH) D levels. Furthermore, This study also found that the interaction between rs7041-GC and rs1562902-CYP2R1, rs10766197-CYP2R1, rs2248137-CYP24A1, rs2242480-CYP34, rs2298849-GC and rs2246709-CYP3A4, the interaction between rs2209314-CYP24A1 and rs1544410-VDR, and plasma 25 (OH) D levels have a significant correlation with the polymorphism interaction of the.GMDR analysis gene. The model of rs1562902-CYP2R1 and rs2209314-CYP2441 is the best model for predicting the level of plasma 25 (OH) D. Conclusion in pregnant women, the polymorphism of vitamin D pathway gene is associated with the level of plasma 25 (OH) D in early pregnancy, and the variation of GC gene may be the major gene variation affecting the level of plasma 25 (OH) D in early pregnancy. There is an interaction between vitamin D metabolic pathway genes and vitamin D levels. The results still need further validation.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.2

【相似文献】