【作者】 刘慧；

【导师】 邵珊珊；

【作者基本信息】 安徽医科大学 ， 公共卫生， 2022， 硕士

【摘要】 目的研究发现孕期妊娠相关焦虑可以预测学龄前儿童的情绪症状和行为发育,并存在性别差异。本研究的目的是在不同队列中验证这种性别差异,并探究性别差异是否与胎盘组织中糖皮质激素反应基因甲基化水平有关。方法本研究依托于马鞍山优生优育队列开展。该队列于2013年5月至2014年9月,在安徽省马鞍山市妇幼保健院招募了3474名基线孕妇,排除不良妊娠结局162例和双胎39例后,纳入3273对单胎活产儿及其母亲,并于产后随访子代生长发育情况。本研究选取其中2405对在孕晚期完成了妊娠相关焦虑评估并在4岁时完成了多动和情绪症状评估的母子对作为研究对象。采用本课题组自编的《孕产期母婴健康记录表》收集孕妇基线资料,使用《妊娠相关焦虑量表》评估孕晚期妊娠相关焦虑情况。由专门人员测量及记录孕妇身高、孕早中晚期血压、血糖和分娩前体重,并于分娩期摘录病案中的出生资料、收集胎盘组织。采用《婴幼儿健康记录表》收集母乳喂养信息,使用《长处与困难问卷(SDQ)》对4岁儿童的多动和情绪症状进行评估。根据《妊娠相关焦虑量表》评价的孕晚期妊娠相关焦虑症状得分排序,选取得分靠前和靠后的各300个胎盘组织进行后续表观遗传学研究。采用Methyl Target法检测胎盘组织中FKBP5、NR3C1和HSD11B2基因启动子区CpGs甲基化水平。探索性因子分析法获得糖皮质激素反应基因甲基化的显著正交因子(简称甲基化因子)。采用Logistic回归模型分析孕晚期妊娠相关焦虑与儿童多动和情绪症状的关联;甲基化因子得分与儿童多动和情绪症状的关联;线性回归模型分析孕晚期妊娠相关焦虑与甲基化因子得分的关联。上述Logistic回归和线性回归分析中均按儿童性别分层分析,并在足月儿和正常出生体重儿亚组中进行敏感性分析。使用PROCESS程序分析甲基化因子得分在孕晚期妊娠相关焦虑与儿童多动和情绪症状关联中的中介作用。结果纳入的2405名孕妇平均年龄为(26.63±3.60)岁。有713人(29.6%)在孕晚期有妊娠相关焦虑,平均年龄为(26.21±3.32)岁。孕晚期无妊娠相关焦虑组与有妊娠相关焦虑组的孕妇在年龄、孕期增重、文化程度、吸烟和分娩方式上的差异具有统计学意义(P<0.05)。在控制混杂因素(孕妇年龄,孕前BMI、孕期增重、文化程度,家庭月收入、吸烟、饮酒、妊娠糖尿病、妊娠高血压、分娩方式和六个月纯母乳喂养)后,与无妊娠相关焦虑组相比,母亲孕晚期妊娠相关焦虑会增加男孩多动的发生风险(OR=1.54,95%CI=1.13～2.11),女孩情绪症状的发生风险(OR=1.61,95%CI=1.15～2.24)。因子分析共提取了五个显著的正交因子(即甲基化因子)。仅在男孩中发现甲基化因子与孕晚期妊娠相关焦虑及学龄前儿童多动和情绪症状的显著关联性,具体为孕晚期妊娠相关焦虑与甲基化因子5得分呈负相关(OR=-0.50,95%CI=-0.87～-0.13),因子5显著负载FKBP5。因子1得分会增加多动症(OR=1.80,95%CI=1.15～2.83)和情绪症状(OR=1.64,95%CI=1.03～2.59)的发生风险,因子4得分会降低情绪症状的发生风险(OR=0.53,95%CI=0.29～0.97)。因子1显著负载NR3C1,因子4显著负载HSD11B2。上述结果经敏感性分析后仍具有显著性。但是,本研究没有发现上述甲基化因子在妊娠相关焦虑与学龄前儿童多动和情绪症状的性别依赖性关联中的中介作用。结论孕晚期妊娠相关焦虑以一种性别依赖的方式预测学龄前儿童多动和情绪症状,这进一步佐证了之前的研究结果。此外,本研究发现胎盘糖皮质激素反应基因甲基化与妊娠相关焦虑和学龄前儿童多动和情绪症状的关联均存在性别依赖性,但并未发现中介作用。原因可能是本研究用于中介效应分析的样本量不够大(所有中介效应的把握度均<80%)。胎盘糖皮质激素反应基因甲基化是否是孕期妊娠相关焦虑致儿童神经行为发育性别差异的表观遗传学机制还需进一步探讨。更多还原

【Abstract】 Objective Studies have found that pregnancy-related anxiety during pregnancy can predict preschoolers’ emotional and behavioral development in a gender-dependent manner.This study aims to test for this gender-specific effect in a different cohort and investigate whether the gender difference was specific to placental methylation of genes regulating glucocorticoids.Methods This study was conducted based on the Ma’anshan Birth Cohort Study(MABC).A total of 3474 baseline pregnant women were recruited in this cohort from May 2013 to September 2014 in Ma’anshan Women’s And Children’s Health Hospital,Anhui Province,After 162 cases of adverse pregnancy outcomes and 39 cases of twins were excluded,3273 pairs of single live births and their mothers were included in the cohort,and the growth and development of their offspring were followed up after delivery.The study included 2,405 mother-child pairs who completed an assessment of pregnancy-related anxiety in the third trimester and mood symptoms and hyperactivity symptoms at 4 years old.Using our research group made "Maternal child health record during pregnancy" pregnant women baseline information collection.The maternal pregnancy-related anxiety symptoms were evaluated with the Pregnancy-Related Anxiety Questionnaire in the third trimester of pregnancy.The height of pregnant women,blood pressure,blood sugar and weight before delivery shall be measured and recorded by specialized personnel,and medical records and birth records shall be extracted during delivery,and placental tissues shall be collected.“The babies and infants health record” was used to collect breastfeeding information,child emotional symptoms and hyperactivity was assessed with the Strengths and Difficulties Questionnaire(SDQ)at 4 years old and the Strengths and Difficulties Questionnaire.According to the score ranking of pregnancy-related anxiety symptoms in the third trimester evaluated by the Pregnancy-related Anxiety Questionnaire,a total of 300 placental tissues with higher and lower scores were selected for subsequent epigenetic studies.Placental methylation of CpGs in promoter regions of FKBP5,NR3C1 and HSD11B2 genes was quantified using the Methyl Target approach.Exploratory factor analysis was used to obtain significant orthogonal factors for the methylation level of glucocorticoid response gene(methylation factor for short).Logistic regression models were used to estimate association of pregnancy-related anxiety in the third trimester with hyperactivity and emotional symptoms,association of methylation factor scores with hyperactivity and mood symptoms in children,a linear regression model was used to analyze the association between trimester pregnancy-related anxiety and methylation factor scores.The above Logistic regression and linear regression analyses were stratified by gender.Sensitivity analysis was performed in full term infants and normal birth weight infants.The mediating role of methylation factor scores between pregnancy-related anxiety in the third trimester and children’s hyperactivity and emotional symptoms was analyzed using the PROCESS procedure.Results The mean age of 2405 pregnant women was(26.63±3.60)years.713(29.6%)had pregnancy-related anxiety in the third trimester,with an average age of(26.21±3.32)years.There were statistically significant differences in maternal age,pregnancy weight gain,education level,smoking and delivery mode between the group without pregnancy related anxiety and the group with pregnancy related-anxiety in the third trimester(P < 0.05).After controlling for confounding factors(maternal age,pre-pregnancy BMI,gestational weight gain,educational level,monthly family monthly income,smoking,drinking,gestational diabetes,pregnancy-induced hypertension,delivery mode,and exclusive breastfeeding in the first 6 months),compared with the group without pregnancy-related anxiety,pregnancy-related anxiety in the third trimester increased the risk of hyperactivity in boys(OR = 1.54,95% CI = 1.13 ～ 2.11)and emotional symptoms in girls(OR = 1.61,95% CI = 1.15 ～2.24).Five significant orthogonal factors were extracted by factor analysis(methylation factor).A significant association between methylation factor and trimester anxiety and hyperactivity and emotional symptoms in preschoolers was found only in boys,with pregnancy-related anxiety in the third trimester negatively associated with methylation factor 5 scores(OR =-0.50,95%CI =-0.87 ～-0.13),and factor 5 was significantly loaded with FKBP5.Factor 1 scores increased the risk of hyperactivity(OR = 1.80,95%CI = 1.15 ～ 2.83)and emotional symptoms(OR = 1.64,95%CI = 1.03 ～ 2.59)in boys,Factor 4 score reduced the risk of emotional symptoms(OR = 0.53,95% CI = 0.29 ～ 0.97).Factor 1 significantly loaded NR3C1 and factor 4significantly loaded HSD11B2.The results were significant after sensitivity analysis.However,no mediating role of methylation factor scores was found in the gender-dependent association between pregnancy-related anxiety and hyperactivity and emotional symptoms in preschoolers.Conclusions Pregnancy-related anxiety in the third trimester predicts hyperactivity and emotional symptoms in preschoolers in a gender-dependent manner,further supporting previous findings.In addition,placental glucocorticoid response gene methylation was found to be gender-dependent in association with pregnancy-related anxiety and hyperactivity and emotional symptoms in preschoolers,but no mediating effect was found.The reason may be that the sample size used for mediating effect analysis in this study was not large enough(the power of all mediating effects was <80%).Whether the methylation of placental glucocorticoid response genes is the epigenetic mechanism of gender differences in neurobehavioral development in children caused by pregnancy-related anxiety during pregnancy remains to be further explored.更多还原