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[目的]观察移行上皮反应基因(TERE1)在小鼠胚胎双前肢正常发育和异常发生中的表达,探讨TERE1在短肢发生中的作用。[方法]ICR小鼠受孕后,将其随机分为实验、对照两组,各64只。于孕第10天(GD10),经口灌胃一次给予实验组孕鼠80 mg/kg的全反式视黄酸(atRA)、对照组孕鼠给予等体积的大豆油,并分别于GD11~GD18取两组胎鼠的双前肢。于GD12取下小鼠胚胎双前肢,以atRA诱导,培养72 h后收获肢。利用实时荧光定量聚合酶链反应方法检测TERE1在各样本中的表达情况。[结果]在整体动物试验中,atRA可诱致小鼠胚胎明显的短肢畸形;在体外试验中,atRA可诱致小鼠胚胎前肢多种骨骼发育异常。TERE1在正常、异常肢及培养肢芽中均有表达。在正常和异常肢中,TERE1的表达模式一致;除GD15外,实验肢中TERE1的表达均低于相同时点正常肢的表达水平。在体外培养肢,各组的表达量随培养时间的延长而增加。实验组在培养24 h,atRA各剂量组的表达量均低于对照组(P<0.05);培养48 h和72 h,在2.5×10-6 mol/L~1.0×10-5 mol/L剂量范围有随剂量增加表达量增加的趋势,但在高剂量组(2.0×10-5 mol/L)表达又下降。[结论]TERE1与小鼠胚胎前肢的发育过程有关,atRA在诱致小鼠前肢异常发生中可引起TERE1 mRNA的表达改变。
[Objective] To investigate the expression of TERE1 gene in the normal development and abnormalities of both forelimbs in mouse embryos and to explore the role of TERE1 in the development of short limbs. [Method] After ICR mice were conceived, they were randomly divided into experimental group and control group, 64 in each. On the 10th day of gestation (GD10), all rats were given orally given 80 mg / kg all-trans retinoic acid (atRA) by oral gavage. The pregnant rats in the control group were given equal volume of soybean oil, GD18 fetus two pairs of fetuses from both forelimbs. The double forelimbs of mouse embryos were removed at GD12, induced by atRA and harvested after 72 h of culture. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the expression of TERE1 in each sample. [Results] atRA could induce obvious short-limb deformity in whole mouse embryos. AtRA could induce various skeletal dysplasia in the mouse forelimbs in in vitro experiments. TERE1 is expressed in normal, abnormal limbs and limb buds. In normal and abnormal limbs, the expression pattern of TERE1 was consistent. Except for GD15, the expression of TERE1 in experimental limbs was lower than that of normal limbs at the same time point. In vitro cultured limbs, the expression of each group increased with the incubation time. At 24 h after culture, the expression of atRA in each dose group was lower than that in the control group (P <0.05). After being cultured for 48 h and 72 h, the expression levels of atRA at doses ranging from 2.5 × 10-6 mol / L to 1.0 × 10-5 mol / L The dose range tends to increase with increasing dose, but decreases again in the high dose group (2.0 × 10-5 mol / L). [Conclusion] TERE1 is related to the development of forelimb in mouse embryos. AtRA induced the expression of TERE1 mRNA in induced forelimb anomalies in mice.