目的 探讨亲代叶酸缺乏对子代大鼠尿嘧啶错误掺入的影响。方法 选用6 w龄雌性Sprague-Dawley（SD）大鼠48只、雄性SD大鼠24只,根据饲料中不同叶酸含量随机分为4组：亲代叶酸均缺乏（parental folate-deficient diet group, D-D）组、母代叶酸缺乏和父代叶酸正常（maternal folate-deficient and paternal folate-normal diet, D-N）组、母代叶酸正常和父代叶酸缺乏（maternal folate-normal and paternal folate-deficient diet, N-D）组、亲代叶酸均正常（parental folate-normal diet, N-N）组,每组12只雌鼠和6只雄鼠。合笼分娩后亲代雌鼠的干预方式不变,子代鼠自断乳后继续饲喂与亲代雌鼠相同的饲料至100日龄（postnatal day, PND）。于PND0、PND50、PND100,采用高效液相色谱法检测子代鼠脑组织中脱氧尿嘧啶（deoxyuridine monophosphate, dUMP）和脱氧胸苷酸（deoxythymidine monophosphate, dTMP）含量。取出生24小时内子代鼠脑海马和纹状体培养神经干细胞（neural stem cells, NSCs）,检测其中dUMP和dTMP含量。结果 D-D组子代鼠脑组织中dTMP含量最低,其次为D-N组、N-D组,N-N组最高;dUMP含量、dUMP/dTMP比值与dTMP相反,D-D组最高,其次为D-N组、N-D组,N-N组最低;dUMP含量及dUMP/dTMP比值均随干预时间增加而升高,dTMP含量随干预时间增加而减少;体外培养的NSCs中四组dTMP含量、dUMP含量、dUMP/dTMP比值与体内实验结果基本一致。结论 亲代叶酸缺乏导致子代大鼠脑组织和体外培养的NSCs中尿嘧啶错误掺入增加,且父代和母代叶酸均缺乏组的影响最大,其次是仅母代叶酸缺乏组,然后是仅父代叶酸缺乏组。
Abstract
Objective To investigate the effect of parental folic acid deficiency on brain uracil misincorporation in offspring rats. Methods Forty-eight 6-week-old female rats and 24 6-week-old male rats were randomly divided into four groups: parental folic acid deficient group (D-D), maternal folic acid deficient and paternal folic acid normal group (D-N), maternal folic acid normal and paternal folic acid deficient group (N-D), and parental folic acid normal group (N-N). Each group had 12 female rats and 6 male rats. The offspring were fed the same diets of their mothers until postnatal day 100. For in vivo study, the deoxythymidine monophosphate (dTMP) and deoxyuridine monophosphate (dUMP) levels in brain tissue of offspring were detected by high-performance liquid chromatography at three different time points. For in vitro study, within 24 h after birth, neural stem cells (NSCs) were cultured for 7 days after being isolated from the hippocampus and striatum, and the dTMP and dUMP levels were measured. Results The level of dTMP in brain tissue was the lowest in the D-D group, followed by the D-N group and the N-D group, and the highest in the N-N group. The changes of dUMP level and dUMP/dTMP ratio were opposite to that of the dTMP. The dUMP level and dUMP/dTMP ratio in the brain tissue of the four groups increased with time, while the dTMP level decreased with time. The changes of dTMP, dUMP and dUMP/dTMP ratio of the NSCs cultured in vitro were consistent with those of brain tissue. Conclusion Parental folic acid deficiency increased uracil misincorporation in the brain tissue of offspring and the NSCs cultured in vitro in which the effect of parental folic acid deficiency was most significant, followed by maternal folic acid deficiency and paternal folic acid deficiency.
关键词
亲代 /
叶酸 /
神经干细胞 /
脱氧胸苷酸 /
脱氧尿嘧啶 /
尿嘧啶错误掺入
Key words
parental /
folic acid /
neural stem cells /
deoxythymidine monophosphate /
deoxyuridine monophosphate /
uracil misincorporation
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基金
国家自然科学基金（No.82003439）; 天津市教委科研计划项目（No.2019KJ166,2022KJ203）
