丙戊酸钠诱导小鼠自闭症模型的改良
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摘要
目的通过改良丙戊酸钠(VPA)小鼠自闭症模型,提高建模成功率,缩短实验周期,更好地模拟临床自闭症发病情况。方法将实验小鼠分为3组,对照组在小鼠孕12.5 d时腹腔注射等体积生理盐水;传统组在小鼠孕12.5 d时一次性腹腔注射VPA600 mg/kg;改良组在小鼠孕10、12 d时,分别腹腔注射VPA 300 mg/kg。观测3组孕鼠给药后反应和6 h死亡率、流产率、临产子宫;观测仔鼠的形态与生长发育;观察仔鼠社会行为学,包括三腔实验、社交实验、青年玩耍实验和旷场实验等反映仔鼠自闭症样行为。结果改良组孕鼠死亡率和流产率低于传统组(P<0.01);与正常组相比,传统组和改良组仔鼠均存在发育障碍(P<0.05),但是改良组新生鼠死亡率明显低于传统组,弯尾高达100%(P<0.001)。与正常组相比,改良组和传统组均出现了自闭症样行为,即社交障碍和重复刻板行为(P<0.05)。结论改良后的给药方法,能更好的模拟人类自闭症发生的过程,同时降低了孕鼠和新生鼠的死亡率、流产率,缩短了实验周期,提高了实验效率。
Objective To establish an improved mouse model of valproic acid(VPA)-induced autism that better mimics human autism. Methods We established mouse models of autism in female C57 mice by intraperitoneal injection of sodium valproate either at a single dose(600 mg/kg) on day 12.5 after conception(conventional group) or in two doses of 300 mg/kg each on days 10 and 12 after conception(modified group), and the control mice were injected with saline only on day 12.5. The responses of the mice to VPA injection, the uterus, mortality rate, and abortion rate were compared among the 3 groups. The morphology and development of the offspring mice were assessed, and their behavioral ontogeny was evaluated using 3-chambered social test, social test, juvenil play test, and open field test. Results The mortality and abortion rates were significantly lower in the modified model group than in the conventional group(P<0.01). Compared with those in the control group, the offspring mice in both the conventional group and the modified group showed developmental disorders(P<0.05).The mortality rate of the newborn mice was significantly lower in the modified group than in the conventional group with a rate of curvy tail of up to 100%(P<0.001). The offspring mice in both the modified group and conventional group exhibited autism-like behavioral abnormalities, including social disorder and repetitive stereotyped behavior(P<0.05). Conclusion The mouse model of autism established using the modified method better mimics human autism with reduced mortality and abortion rates of the pregnant mice and also decreased mortality rate of the newborn mice.
Objective To establish an improved mouse model of valproic acid(VPA)-induced autism that better mimics human autism. Methods We established mouse models of autism in female C57 mice by intraperitoneal injection of sodium valproate either at a single dose(600 mg/kg) on day 12.5 after conception(conventional group) or in two doses of 300 mg/kg each on days 10 and 12 after conception(modified group), and the control mice were injected with saline only on day 12.5. The responses of the mice to VPA injection, the uterus, mortality rate, and abortion rate were compared among the 3 groups. The morphology and development of the offspring mice were assessed, and their behavioral ontogeny was evaluated using 3-chambered social test, social test, juvenil play test, and open field test. Results The mortality and abortion rates were significantly lower in the modified model group than in the conventional group(P<0.01). Compared with those in the control group, the offspring mice in both the conventional group and the modified group showed developmental disorders(P<0.05).The mortality rate of the newborn mice was significantly lower in the modified group than in the conventional group with a rate of curvy tail of up to 100%(P<0.001). The offspring mice in both the modified group and conventional group exhibited autism-like behavioral abnormalities, including social disorder and repetitive stereotyped behavior(P<0.05). Conclusion The mouse model of autism established using the modified method better mimics human autism with reduced mortality and abortion rates of the pregnant mice and also decreased mortality rate of the newborn mice.
引文
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[15]Yang EJ,Ahn S,Lee K,et al.Early behavioral abnormalities and perinatal alterations of PTEN/AKT pathway in valproic acid autism model mice[J].PLoS One,2016,11(4):298-307.
[16]Meador K,Reynolds MW,Crean S,et al.Pregnancy outcomes in women with epilepsy:A systematic review and meta-analysis of published pregnancy registries and cohorts[J].Epilepsy Res,2008,81(1):1-13.
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[18]Meador KJ,Baker GA,Browning N,et al.Foetal antiepileptic drug exposure and verbal versus non-verbal abilities at three years of age[J].Brain,2011,134(2):396-408.
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[23]雷雨溪,何慕兰,赖茜,等.两种孤独症谱系障碍大鼠模型的构建及比较[J].重庆医科大学学报,2018,43(9):1132-9.
[24]Sato M,Shirota M,Nagao T.Pantothenic acid decreases valproic acid-induced neural tube defects in mice[J].Teratology,1995,52(3):143-8.
[25]Al-Amin MM,Rahman MM,Khan FR,et al.Astaxanthin improves behavioral disorder and oxidative stress in prenatal valproic acid-induced mice model of autism[J].Behav Brain Res,2015,286(2):112-21.
[26]Mony TJ,Lee JW,Dreyfus CA,et al.Valproic acid exposure during early postnatal gliogenesis leads to autistic-like behaviors in rats[J].Clin Psychopharmacol Neuros,2016,14(4):338-44.
[27]Hill DS,Robert C,Deeann WS,et al.Autism-Like behavior and epigenetic changes associated with autism as consequences of\r,in utero\r,exposure to environmental pollutants in a mouse model[J].Behav Neurol,2015,20(1):1-10.
[28]Coretti L,Cristiano C,Florio E,et al.Sex-related alterations of gut microbiota composition in the BTBR mouse model of autism spectrum disorder[J].Sci Rep,2017,35(7):45356-67.
[29]侯倩伶.VPA孤独症大鼠发育特点和GABA能神经元在不同脑区的表达[D].重庆:学重庆医科大学,2017.
[30]Rasalam AD,Hailey H,Williams JH,et al.Characteristics of fetal anticonvulsant syndrome associated autistic disorder[J].Dev Med Child Neurol,2005,47(8):551-5.
[2]Ioana CA,Stefanachi EA.Autism,an overwhelming condition:history,etiopathogenesis,types,diagnosis,therapy and prognosis[J].Rev Med Chir Soc Med Nat Iasi,2013,117(3):654-61.
[3]Gray K,Keating C,Taffe J,et al.Trajectory of behavior and emotional problems in autism[J].Am J Intellect Dev Disabil,2012,117(2):121-33.
[4]Das N,Dhanawat M,Shrivastava SK.An overview on antiepileptic drugs[J].Drug Discov Ther,2012,6(4):178-93.
[5]Mohiuddin S,Ghaziuddin M.Psychopharmacology of autism spectrum disorders:a selective review[J].Autism,2013,17(6):645-54.
[6]Gomes PM,Lima LL,Bueno MG,et al.Autism in Brazil:a systematic review of family challenges and coping strategies[J].JPediatr(Rio J),2015,91(2):111-21.
[7]Parish S,Thomas K,Rose R,et al.State insurance parity legislation for autism services and family financial burden[J].Intellect Dev Disabil,2012,50(3):190-8.
[8]Schneider T,Przew?ocki R.Behavioral alterations in rats prenatally exposed to valproic acid:animal model of autism[J].Neuropsychopharmacology,2005,30(1):80-9.
[9]Singer HS,Morris C,Gause C,et al.Prenatal exposure to antibodies from mothers of children with autism produces neurobehavioral alterations:a pregnant dam mouse model[J].J Neuroimmunol,2009,211(1/2):39-48.
[10]Shultz SR,Macfabe DF,Martin SA,et al.Intracerebroventricular injections of the enteric bacterial metabolic product propionic acid impair cognition and sensorimotor ability in the long-evans rat:further development of a rodent model of autism[J].Behav Brain Res,2009,200(1):33-41.
[11]WinterC,ReutimanTJ,Folsom TD,et al.Dopamine and seroton in levels following prenatal viral in fection in mouse-implications for psychiatric disorders such as schizophrenia and autism[J].Europ Neuropsychopharmacol,2008,18(10):712-6.
[12]Moy SS,Nonneman RJ,Young NB,et al.Impaired sociability and cognitive function in Nrcam-null mice[J].Behav Brain Res,2009,205(1):0-131.
[13]Choi CS,Gonzales EL,Kim KC,et al.The transgenerational inheritance of autism-like phenotypes in mice exposed to valproic acid during pregnancy[J].Sci Rep,2016,6(12):36250-67.
[14]Hou QL,Yan W,Li YB,et al.A developmental study of abnormal behaviors and altered GABAergic signaling in the VPA-treated rat model of Autism[J].Front Behav Neurosci,2018,(12):182-91.
[15]Yang EJ,Ahn S,Lee K,et al.Early behavioral abnormalities and perinatal alterations of PTEN/AKT pathway in valproic acid autism model mice[J].PLoS One,2016,11(4):298-307.
[16]Meador K,Reynolds MW,Crean S,et al.Pregnancy outcomes in women with epilepsy:A systematic review and meta-analysis of published pregnancy registries and cohorts[J].Epilepsy Res,2008,81(1):1-13.
[17]Meador KJ,Baker GA,Browning NA,et al.Cognitive function at3 years of age after fetal exposure to antiepileptic drugs[J].N Engl J Med,2009,360(16):1597-605.
[18]Meador KJ,Baker GA,Browning N,et al.Foetal antiepileptic drug exposure and verbal versus non-verbal abilities at three years of age[J].Brain,2011,134(2):396-408.
[19]Nadebaum C,Anderson V,Vajda F,et al.The Australian brain and cognition and antiepileptic drugs study:IQ in school-aged children exposed to sodium valproate and polytherapy[J].J Intern Neuropsychol Society,2011,17(1):133-42.
[20]Shallcross R,Bromley RL,Irwin B,et al.Child development following in utero exposure:levetiracetam vs sodium valproate[J].Neurology,2011,76(4):383-9.
[21]李天苏,潘娜娜,杨李,等.Sprague-Dawley大鼠孤独症模型的建立及行为学特点分析[J].安徽医学,2013,34(7):865-8.
[22]Escalona-Cardoso GN,Paniagua-Castro N,Pérez-Pastén R,et al.Spirulina(arthrospira)protects against valproic Acid-Induced neural tube defects in imprinting control region mice[J].J Med Food,2012,15(12):1103-8.
[23]雷雨溪,何慕兰,赖茜,等.两种孤独症谱系障碍大鼠模型的构建及比较[J].重庆医科大学学报,2018,43(9):1132-9.
[24]Sato M,Shirota M,Nagao T.Pantothenic acid decreases valproic acid-induced neural tube defects in mice[J].Teratology,1995,52(3):143-8.
[25]Al-Amin MM,Rahman MM,Khan FR,et al.Astaxanthin improves behavioral disorder and oxidative stress in prenatal valproic acid-induced mice model of autism[J].Behav Brain Res,2015,286(2):112-21.
[26]Mony TJ,Lee JW,Dreyfus CA,et al.Valproic acid exposure during early postnatal gliogenesis leads to autistic-like behaviors in rats[J].Clin Psychopharmacol Neuros,2016,14(4):338-44.
[27]Hill DS,Robert C,Deeann WS,et al.Autism-Like behavior and epigenetic changes associated with autism as consequences of\r,in utero\r,exposure to environmental pollutants in a mouse model[J].Behav Neurol,2015,20(1):1-10.
[28]Coretti L,Cristiano C,Florio E,et al.Sex-related alterations of gut microbiota composition in the BTBR mouse model of autism spectrum disorder[J].Sci Rep,2017,35(7):45356-67.
[29]侯倩伶.VPA孤独症大鼠发育特点和GABA能神经元在不同脑区的表达[D].重庆:学重庆医科大学,2017.
[30]Rasalam AD,Hailey H,Williams JH,et al.Characteristics of fetal anticonvulsant syndrome associated autistic disorder[J].Dev Med Child Neurol,2005,47(8):551-5.