五子衍宗丸对NTDs胎鼠神经管细胞凋亡的影响

Effect of Wuzi Yanzong Pills on Apoptosis of Neural Tube Cells in NTDs Model

目的:研究五子衍宗丸对全反式维甲酸(all-trans-retinoic acid,ATRA)诱导的神经管畸形(neural tube defects,NTDs)胎鼠模型神经管细胞凋亡的影响。方法:将BALB/c小鼠以雌雄2∶1比例合笼,阴栓阳性者为孕鼠。将孕鼠随机分为正常组、模型组、五子衍宗丸先给组、五子衍宗丸和ATRA同时干预组、五子衍宗丸后给组,后3组又各分为高、中、低剂量3个亚组(4.20,2.10,1.05 g·kg-1)。在胚胎7 d时,模型组、五子衍宗丸先给组的3个亚组、五子衍宗丸和ATRA同时干预组的3个亚组、五子衍宗丸后给组的3个亚组均给孕鼠灌胃50 mg·kg-1ATRA 1次。在胚胎11 d时,处死孕鼠取出胚胎,制成横切片后进行HE染色和TUNEL染色,观察各组神经管发育情况和计算神经管细胞凋亡指数。结果:光镜下观察HE染色结果,正常组神经管发育良好,闭合完全;模型组神经管发育不良,闭合不全;五子衍宗丸中、高剂量先给组,五子衍宗丸中、高剂量和ATRA同时干预组神经管发育较好,管壁可见完整的神经上皮层、套层、边缘层,神经上皮细胞排列较致密有序;五子衍宗丸中、高剂量后给组和五子衍宗丸低剂量各组神经管发育不良,神经上皮细胞数量减少、排列无序。光镜下观察TUNEL染色结果,与正常组比较,五子衍宗丸高剂量先给组凋亡指数的差异无统计学意义;其余各组凋亡指数增高的差异均有统计学意义。与模型组比较,五子衍宗丸中、高剂量先给组,五子衍宗丸中、高剂量和ATRA同时干预组凋亡指数显著降低,差异均有统计学意义;五子衍宗丸中、高剂量后给组和五子衍宗丸低剂量各组凋亡指数的差异均无统计学意义。结论:五子衍宗丸可抑制NTDs胎鼠模型神经管细胞的凋亡。

Objective：To study the influence of Wuzi Yanzong pills on apoptosis of neural tube cells in neural tube defects （NTDs） model induced by all-trans-retinoic acid （ATRA）.Method：BALB/c females and males （ratio 2∶1） were caged together overnight.Checked for the presence of a vaginal plug,the females were diagnosed as pregnant mice.Pregnant mice were randomly divided into normal group,model group,the group of early administration of Wuzi Yanzong pills,the group of simultaneous administration of Wuzi Yanzong pills and ATRA,and the group of late administration of Wuzi Yanzong pills.Each Wuzi Yanzong pills group was divided into three subgroups of high,middle and low doses （4.20,2.10,1.05 g ·kg^-1）.Model group and three subgroups of group of early administration of Wuzi Yanzong pills and group of simultaneous administration of Wuzi Yanzong pills and ATRA and group of late administration of Wuzi Yanzong pills were once fed with 50 mg ·kg^-1ATRA at embryo 7 d.After pregnant mice were sacrificed at embryo 11 d,embryos were separated.The embryos were performed by preparation for sections using the staining of HE and TUNEL,which were used to observe the development of neural tube and calculate apoptotic index.Result：The neural tube developed well and completely closed in normal group.The neural tube appeared dysplasia in model group.The neural tube developed well,and consisted of ependymal layer,mantle layer and marginal layer,and neuroepithelial cells arranged in order in groups of early administration of middle and high dose Wuzi Yanzong pills and groups of simultaneous administration of middle and high dose Wuzi Yanzong pills and ATRA.The neural tube appeared dysplasia,and neuroepithelial cells reduced and arranged in disorder in groups of late administration of middle and high dose Wuzi Yanzong pills and the groups of low dose Wuzi Yanzong pills.Compared with the normal group,the apoptotic index in the group of early administration of high dose Wuzi Yanzong pills have no statistically significant difference,but the apoptotic index in other groups were significantlyincreased.Compared with the model group,the apoptotic index in the groups of early administration of middle and high dose Wuzi Yanzong pills and the groups of simultaneous administration of middle and high dose Wuzi Yanzong pills and ATRA were significantly decreased; but the apoptotic index in the groups of late administration of middle and high dose Wuzi Yanzong pills and the groups of low dose Wuzi Yanzong pills showed no statistically significant decline.Conclusion：Wuzi Yanzong pills can inhibit the apoptosis of neural tube cells in NTDs model.