丙戊酸诱导胎鼠发生隐性脊椎裂畸形的模型建立

Model of spina bif ida occulta induced by valproic acid in foetal rats

摘要:

目的 研究丙戊酸对Wistar大鼠的致畸作用。方法 2 4只雌性受孕Wistar大鼠随机分成五组 ,不同剂量丙戊酸 (4 0 0mg/kg ,4 5 0mg/kg ,5 0 0mg/kg)三组、生理盐水对照组和空白对照组 ,丙戊酸的三组在妊娠第9天 9am和 4am皮下注射丙戊酸钠溶液两次 ,妊娠第 2 0天剖宫产取出胚胎 ,脊柱标本进行骨和软骨双重染色后进行显微图象分析 ,测量椎体椎弓两个软骨端的距离并与正常范围值比较 ,判定隐性脊椎裂。结果 Wistar大鼠正常 2 0天胚胎 ,胸 9到骶 3椎体椎弓两个软骨端的距离正常值范围小于 1 6 6 .4 μm(1 1 2 .0± 1 .96× 2 7.7μm上限 ) ,骶4椎体正常值小于 1 85 .5 μm(1 2 7.7± 1 .96× 2 9.5 μm 上限 ) ,骶 5椎体正常值小于 1 98.7μm(1 4 2 .1± 1 .96×2 8.9μm上限 )。盐水对照组胚胎无畸形发生 ;丙戊酸钠 4 0 0mg/kg组 ,隐性脊椎裂发生率 80 % ;丙戊酸钠 4 5 0mg/kg组发生率达 93% ;丙戊酸钠 5 0 0mg/kg组发生率达到 1 0 0 %。其中一个胚胎出现头面部、颅骨、脊柱、尾多个部位畸形。结论 丙戊酸能诱导胎鼠发生隐性脊椎裂畸形

Abstract:

Objective To study the teratogenicity of valproic acid (vpa) in Wistar rats. Methods Twenty four pregnant Wistar rats were randomly assigned into 5 groups: 3 VPA of different dosages groups (400 mg/kg, 450 mg/kg and 500 mg/kg groups), the normal saline (NS) control group and the Blank control group. On the 9th day of gestation, 3 VPA groups were subcutaneously injected different dosages of vpa at 9am and 4pm. The NS control group was administrated 1 ml of NS instead and the Blank control group had no intervention. On the 20th day of gestation, the embryos were taken out and the specimens from spines were double stained with Alcian blue GX and Alizarin red S. The distance of the two cartilaginous ends of the vertebra arch was measured with a stereomicroscope and compared with the normal limit. Results In the Blank control group, the distance of the two vertebral cartilaginous ends was less than 166.4 μm (the superior limit of 112.0±1.96×27.7 μm) from the 9th thoracic to the 3rd sacral vertebra; less than 185.5 μm (the superior limit of 127.7±1.96×29.5 μm) in the 4th sacral vertebra; and less than 198.7 μm (the superior limit of (142.1±1.96×28.9 μm) in the 5th sacral vertebra. There was no malformation in the NS control group. The incidence of spina bifida occulta was 80%, 93% and 100% in the VPA 400 mg/kg, 450 mg/kg and 500 mg/kg groups respectively. The craniofacial, skull, spine and tail malformations occurred in a fetus in the vpa 500 mg/kg group. Conclusions Vpa could induce spina bifida occulta in foetal rats.