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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 investigate the effect of Wuzi Yanzong Pills on apoptosis of neural tube cells in a fetal rat model of neural tube defects (NTDs) induced by all-trans-retinoic acid (ATRA). Methods: The BALB / c mice were co-housed in a ratio of 2: 1, and the positive ones were pregnant rats. The pregnant rats were randomly divided into normal group, model group, Wuzi Yanzong pill first group, Wuzi Yanzong pill and ATRA intervention group, Wuzi Yanzong Pills group, the latter three groups were divided into high, medium and low dose three Subgroups (4.20, 2.10, 1.05 g · kg -1). At embryonic day 7, three subgroups of model group, Wuzi Yanzong pill first group, Wuzi Yanzong pill and ATRA group, three subgroups of Wuziyanzong pill group, Oral stomach 50 mg · kg-1ATRA 1 times. On the 11th day of embryo, the pregnant mice were sacrificed and the embryos were removed and made into transverse sections for HE staining and TUNEL staining. The development of neural tube in each group and the apoptosis index of neural tube were calculated. Results: Under the light microscope, the results of HE staining showed that the neural tube in the normal group developed well and completely closed; the neural tube in the model group was dysplasia and closed incompletely; the middle and high dose Wuziyanzong pill group, Wuziyanzong pill medium and high dose and ATRA At the same time, the development of neural tube in the intervention group was better. The intact neuroepithelium, the lamina, the marginal layer and the neuroepithelial cells in the intervention group were arranged more densely and orderly. The middle and high doses of Wuzi Yanzong Pill and the low doses of Wuzi Yanzong Pill Group of neural tube dysplasia, reduce the number of neuroepithelial cells, arranged in disorder. The results of TUNEL staining were observed under light microscope. There was no significant difference in apoptotic index of Wiziyanzong pill high-dose pre-treatment group compared with normal group. The difference of apoptosis index in the other groups was statistically significant. Compared with the model group, Wuziyanzong pill medium and high doses of the first group, Wuzi Yanzong Pills medium and high doses and ATRA simultaneous intervention group significantly reduced the apoptotic index, the differences were statistically significant; Group A and Wuzi Yanzong Pills in low dose groups showed no significant difference in apoptosis index. Conclusion: Wuzi Yanzong pill can inhibit the apoptosis of neural tube cells in NTDs fetal rat model.