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目的:通过检测WIF-1和SFRP-1在环磷酰胺(CP)致大鼠神经管畸形(NTDs)中神经上皮细胞的表达及表达的变化,研究CP对胎鼠神经上皮细胞Wnt信号通路的影响,探讨NTDs发生的分子机制,为NTDs的预防、基因治疗提供理论依据。方法:将SD孕鼠随机分为实验组和对照组,分别于妊娠13天8～9时一次性腹腔注射CP(15 mg/kg)和等量的生理盐水。分别于给药后8、12、24 h处死孕鼠,剖腹取出胎鼠,用10%中性福尔马林固定,常规石蜡包埋、连续切片。应用免疫组织化学方法检测WIF-1与SFRP-1在神经管神经上皮细胞的表达及其变化。结果:①WIF-1免疫组化显示,给药后8 h,实验组WIF-1阳性细胞分布与对照组相似(P>0.05);给药后12 h,实验组WIF-1阳性表达增强(P<0.05);给药后24 h,实验组与对照组相比,神经管背侧、顶板区WIF-1阳性表达明显增强(P<0.01)。②SFRP-1免疫组化显示,SFRP-1在实验组和对照组各个时间段均有阳性表达,但差异无统计学意义(P>0.05)。结论:①WIF-1在神经上皮细胞表达增强与CP致NTDs密切相关。②SFRP-1在神经上皮细胞表达的改变未发现与CP致NTDs有相关性。③Wnt信号通路异常与CP致NTDs密切相关。
OBJECTIVE: To investigate the expression of WIF-1 and SFRP-1 in neural tube defects (NTDs) induced by cyclophosphamide (CP) in rats and the changes of Wnt signaling pathway in fetal rat neural epithelial cells Influence, explore the molecular mechanism of NTDs, provide a theoretical basis for the prevention of NTDs, gene therapy. Methods: Pregnant SD rats were randomly divided into experimental group and control group. CP (15 mg / kg) and normal saline (CP) were intraperitoneally injected at 8-9 days of gestation on the 13th day. Pregnant mice were sacrificed at 8, 12, and 24 h after dosing respectively. Fetal rats were removed by cesarean section, fixed in 10% neutral formalin, embedded in paraffin and continuously sliced. Immunohistochemistry was used to detect the expression of WIF-1 and SFRP-1 in neural tube epithelial cells and their changes. Results: ①WIF-1 immunohistochemistry showed that the WIF-1 positive cells in the experimental group were similar to the control group at 8 h after administration (P> 0.05), and the positive expression of WIF-1 in the experimental group was increased 12 h after the administration <0.05). Compared with the control group, the expression of WIF-1 in the dorsal and apical region of the neural tube in experimental group was significantly increased 24 h after administration (P <0.01). (2) SFRP-1 immunohistochemistry showed that SFRP-1 was positively expressed in both experimental and control groups at various time points, but the difference was not statistically significant (P> 0.05). Conclusion: ① The expression of WIF-1 in neuroepithelial cells is closely related to CP induced NTDs. ② The changes of SFRP-1 in neuroepithelial cells were not found to be correlated with CP-induced NTDs. Wnt signaling pathway abnormalities and CP-induced NTDs are closely related.