miR-200b参与2,3,7,8-四氯二苯并二噁英诱导的小鼠发生腭裂

来源 :2016(第二届)毒性测试替代方法与转化毒理学(国际)学术研讨会暨有害结局路径(AOP)与风险评估培训会议 | 被引量 : 0次 | 上传用户:dongfan1909
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  目的 观察miR-200b 基因在 2,3,7,8-四氯二苯并二噁英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)诱导小鼠腭裂实验组和对照组发育过程中的表达变化,探讨miR-200b基因在TCDD诱导小鼠腭裂发生中的作用机制.方法 将50只孕鼠随机分为实验组与对照组.在小鼠孕第10天(gestation day 10,GD10),实验组以60μg/kg TCDD灌胃,对照组以相同途径给与等体积玉米油.两组不同胎龄各取5只母鼠,于GD 12.5、GD 13.5、GD 14.5、GD 15.5和GD 16.5时,用乙醚进行麻醉并处死孕鼠,剖腹取出胎鼠,一部分胎鼠头部经石蜡包埋、HE染色制作组织切片,光学显微镜下观察胎鼠腭裂的发生率.另一部分胎鼠取GD 12.5、GD 13.5、GD 14.5、GD 15.5和GD 16.5时的胎鼠腭突组织,并提取RNA.实时定量PCR检测腭突组织中miR-200b基因的表达情况. 结果 实验组胎鼠全部发生腭裂(100%),对照组胎鼠均未发生腭裂.实时定量PCR结果显示在胎鼠腭突发育的关键时期,GD 12.5实验组miR-200b表达量是对照组的(0.65±0.10)(P<0.05)倍;GD 13.5实验组miR-200b表达量是对照组的(1.71±0.16)倍(P<0.05); GD 14.5实验组miR-200b表达量是对照组的(2.29±0.49)倍(P<0.05); GD 15.5实验组miR-200b表达量是对照组的(1.11±0.10)倍(P<0.05); GD 16.5实验组miR-200b表达量是对照组的(0.66±0.37)倍.结论 60ug/kg TCDD灌胃可以诱导胎鼠发生稳定腭裂.在腭突发育的关键时期,实验组miR-200b基因在TCDD诱导小鼠腭裂发育的关键时期(GD 13.5、GD 14.5和GD 15.5)时表达量升高,miR-200b基因可能参与TCDD诱导腭裂的发生.
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