基于脑-肠轴理论研究氟化物的摄入对于VPA小鼠孤独症样行为的影响及机制  

Effects and mechanisms of fluoride intake on autism-like behaviour in VPA mice based on brain-gut axis theory

作　　者：段娟 辛金鸽 彭薇淇 黄咏梅 周俊球 DUAN Juan;XIN Jin-ge;PENG Wei-qi;HUANG Yong-mei;ZHOU Jun-qiu(Prenatal Diagnostic Center,Baiyun District Maternal and Child Health Hospital,Guangzhou,510500,Guangdong,China;Microecology Research Center,Baiyun Branch,Nanfang Hospital,Southern Medical University,Guangzhou,510500,Guangdong,China;School of Animal Medicine,Sichuan Agricultural University,Chengdu 611130,Sichuan,China)

机构地区：[1]广州市白云区妇幼保健院产前诊断中心,510500 [2]南方医科大学南方医院白云分院微生态研究中心,510500 [3]四川农业大学动物医学院,611130

出　　处：《现代消化及介入诊疗》2023年第12期1505-1513,共9页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：广东省基础与应用基础研究基金项目(2021A1515220009);广州市科技计划项目(202002030186);西藏自治区科技计划项目(XZ202201ZY0027G)。

摘　　要：目的 揭示氟化物的摄入是否可以加剧丙戊酸(VPA)小鼠的孤独症样行为表现,并探索造成这一现象的机制和作用途径。方法 动物试验分为两次进行:第一次试验将3周龄C57雄性小鼠随机分为对照组(Control组)、丙戊酸处理组(VPA组)、低浓度氟化物组(F1组)、中浓度氟化物组(F2组)和高浓度氟化物组(F3组),每组12只,在小鼠7周龄时进行埋藏食物实验、三箱社交实验、旷场实验、埋珠实验以及高架十字迷宫相关行为学检测。测定大脑皮层NR2A、NR2B、BDNF、GABRA1、GAD65和GAD67的mRNA表达和蛋白水平,并进行右旋糖酐法以及DAO和D-乳酸活性检测肠道通透性。为验证迷走神经在氟化物影响下的参与作用,第二次试验将3周龄C57雄性小鼠随机分为VPA伪切断(SC组)、VPA切断组(VC组)、VPA伪切断+F(SF组)、VPA切断+F(VF组),在小鼠7周龄时进行埋藏食物实验、三箱社交实验、旷场实验相关行为学检测。结果VPA组小鼠的孤独症样行为较Control组小鼠更为显著;中、高浓度的氟化物摄入使VPA小鼠相关因子的蛋白水平和mRNA水平均显著降低;肠道通透性检测结果显示,中、高浓度的氟化物摄入导致VPA小鼠肠道高通透性和血清DAO含量和d-乳酸活性的增加;氟化物摄入引起的迷走神经刺激可能在VPA小鼠ASD样行为的表达中发挥作用。结论 VPA小鼠具有显著的ASD行为异常;中、高浓度的氟化物摄入会显著增加VPA小鼠的ASD样行为;这一改变不仅与肠道通透性的改变有关,也与腹部迷走神经的刺激有关。Objective To reveal whether fluoride intake can exacerbate autism-like behavioural manifestations in valproic acid(VPA) mice and to explore the mechanisms and pathways of action responsible for this phenomenon.Methods Animal experiments were conducted twice:in the first experiment,3-week-old C57 male mice were randomly divided into a control group(Control group),a valproic acid-treated group(VPA group),a low-concentration fluoride group(F1 group),a medium-concentration fluoride group(F2 group),and a high-concentration fluoride group(F3 group),with 12 in each group,and were subjected to the buried-food experiment,the three-box socialisation experiment,the open-field experiment,the buried bead experiment,and the elevated cross maze-related behavioural tests when the mice were 7 weeks old.,buried bead experiment and elevated cross maze related behavioural tests.The mRNA expression and protein levels of NR2A,NR2B,BDNF,GABRA1,GAD65,and GAD67 in the cerebral cortex were determined,and intestinal permeability was measured by the dextrose method as well as by DAO and D-lactate activities.To verify the involvement of the vagus nerve under the influence of fluoride,the second experiment randomly divided 3-week-old C57 male mice into VPA pseudo-cut-off(SC group),VPA cut-off group(VC group),VPA pseudo-cut-off+F(SF group),and VPA cut-off+F(VF group),and performed behavioural assays related to the buried food experiment,three-box socialization experiment,and absenteeism experiment when the mice were 7 weeks old.Results The autism-like behaviour of mice in the VPA group was more pronounced than that of mice in the Control group;the protein levels and mRNA levels of the relevant factors in VPA mice were significantly reduced by fluoride ingestion at medium and high concentrations;the results of intestinal permeability assay showed that fluoride ingestion at medium and high concentrations resulted in intestinal hyperpermeability and an increase in serum DAO content and d-lactic acid activity in the VPA mice;and vagal stimulation

关 键 词：氟化物摄入 VPA小鼠 孤独症样行为 迷走神经 

分 类 号：R57[医药卫生—消化系统]