围妊娠期高血清叶酸水平雌性小鼠模型的建立及高血清叶酸水平对胚胎心脏发育的影响  

Establishment of a female mouse model with high serum folate levels during peri-pregnancy and effects of high serum folate levels on embryonic heart development

作　　者：郭姝怡 高娟 王瀚宇 董家豪 谢岩 胡斯琦 安心宇 李桐帆 薛赓 GUO Shu-yi;GAO Juan;WANG Han-yu;DONG Jia-hao;XIE Yan;HU Si-qi;AN Xin-yu;LI Tong-fan;XUE Geng(Department of Medical Genetics,College of Basic Medical Sciences,Naval Medical University(Second Military Medical University),Shanghai 200433,China;Department of Recuperation(Ⅴ),Qingdao Special Servicemen Recuperation Center of PLA Navy,Qingdao 266071,Shandong,China;The Fourth Student Team,College of Basic Medical Sciences,Naval Medical University(Second Military Medical University),Shanghai 200433,China)

机构地区：[1]海军军医大学(第二军医大学)基础医学院医学遗传学教研室,上海200433 [2]海军青岛特勤疗养中心疗养五科,青岛266071 [3]海军军医大学(第二军医大学)基础医学院学员四大队,上海200433

出　　处：《海军军医大学学报》2023年第6期666-671,共6页Academic Journal of Naval Medical University

基　　金：国家自然科学基金(81971402);国家重点研发计划(2016YFC1000503);海军军医大学(第二军医大学)青年启动基金(2022QN010)。

摘　　要：目的通过高叶酸含量配方饲料结合血清叶酸检测建立围妊娠期高血清叶酸水平的雌性小鼠模型,并研究围妊娠期增补叶酸导致血清叶酸水平升高对后代心脏发育的影响。方法根据文献报道设计了正常叶酸含量(2.0 mg/kg)和高叶酸含量(10.0、20.0、40.0 mg/kg)的配方饲料。30只6周龄雌性BALB/c小鼠随机分成5组(每组6只),其中1组使用普通饲料喂养,其余4组分别使用上述4种配方饲料喂养。喂养2周后,根据血清叶酸水平对4组配方饲料喂养的雌鼠(24只)重新分组:正常血清叶酸组(血清叶酸水平<2000 pg/mL)、高血清叶酸组(2000 pg/mL≤血清叶酸水平<3000 pg/mL)、超高血清叶酸组(血清叶酸水平≥3000 pg/mL)。分组后继续使用相应的配方饲料饲养,并与普通饲料喂养的雄鼠合笼,以发现阴道栓当天作为孕0.5 d(E0.5)计算孕龄。获取不同孕龄的胎鼠心脏组织样本,通过H-E染色、免疫组织化学染色对孕10.5 d(E10.5)、孕11.5 d(E11.5)和孕13.5 d(E13.5)胎鼠心脏发育相关指标进行检测。结果随着饲料中叶酸含量的增加,雌鼠的平均血清叶酸水平呈现逐渐升高的趋势,但是不同组间血清叶酸水平差异没有统计学意义(P=0.163)。相较于正常血清叶酸组和高血清叶酸组,超高血清叶酸组胎鼠的心脏表现出间叶组织缺乏(E10.5)、心肌细胞减少且增生不活跃(E10.5)、心房与心室壁厚度均减少(E11.5,P均<0.001)、磷酸化组蛋白H3阳性率降低(E11.5,P均<0.001)、背侧间充质突出缺失(E13.5)、右心耳和心室流出道结构异常(E13.5)及原发孔型房间隔缺损等房室间隔缺损表现(E13.5)。结论通过围妊娠期增补叶酸结合血清叶酸检测构建高血清叶酸水平雌性小鼠模型的方法可行、有效。围妊娠期雌性小鼠增补叶酸导致的血清叶酸水平剧烈升高与后代心脏发育异常相关。Objective To establish a female mouse model of high serum folate levels during peri-pregnancy by feeding a formula feed with high folate contents combined with serum folate testing,and to study the effects of high folate levels caused by folate supplementation during peri-pregnancy on offspring heart development.Methods According to the literatures,the formula feeds with normal folate content(2.0 mg/kg)and high folate content(10.0,20.0,and 40.0 mg/kg)were designed.Thirty 6-week-old female BALB/c mice were randomly divided into 5 groups(6 in each group),1 of which continued to be fed with common feed,and the other 4 groups were fed with the above 4 kinds of formula feeds.After 2 weeks of feeding,4 groups of mice fed with formula feeds were divided into normal serum folate group(serum folate level<2000 pg/mL),high serum folate group(2000 pg/mL≤serum folate level<3000 pg/mL),and ultra-high serum folate group(serum folate level≥3000 pg/mL).After grouping,the female mice continued to be fed with the corresponding formula feeds and caged with male mice feeding with common feed.The gestational age was calculated as 0.5 d of pregnancy(E0.5)on the day when the vaginal plug was found.Cardiac tissue samples from fetal mice of different gestational ages were obtained,and cardiac development indicators at 10.5,11.5 and 13.5 d of pregnancy(E10.5,E11.5,and E13.5)were detected by hematoxylin-eosin and immunohistochemical staining.Results With the increase of folate content in feed,the average serum folate levels of the female mice presented a gradually increasing trend,but there was no significant difference in serum folate level among different groups(P=0.163).Compared to the normal and high serum folate groups,in the ultra-high serum folate group the cardiac tissue of fetal mice showed a loss of mesenchymal tissue(E10.5),decreases in the number and proliferation of myocardial cells(E10.5),thinner atrial and ventricular walls(E11.5,all P<0.001),a decrease in the phosphorylated histone H3 positive rate(E11.5,both P<0.001),a

关 键 词：围妊娠期 增补叶酸 血清叶酸水平 心脏发育 动物模型 

分 类 号：R715.3[医药卫生—妇产科学] R977.22[医药卫生—临床医学]