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目的探讨灵芝孢子能否促进受维甲酸诱导,而停滞在G0/G1期的胚胎神经管神经上皮细胞重新进入细胞周期循环,继续增殖和分化,减少神经管畸形的发生。方法于实验对照组和灵芝孢子组小鼠孕期胚胎(E)7.75d时一次性胃饲全反式维甲酸,造成这两组孕鼠的胚胎出现神经管畸形。然后,给予灵芝孢子组孕鼠胃饲灵芝孢子溶液。在孕期E10.5d时取出两组孕鼠的胚胎,分别计数胚胎的神经管畸形发生率。应用免疫荧光组织化学染色和流式细胞仪检测胚胎神经管神经上皮的巢蛋白(nestin)表达情况。应用DNA定量荧光染色和RT-PCR技术检测胚胎神经管依赖细胞周期蛋白激酶2(Cdk2)mRNA和Cdk4 mRNA的转录。结果灵芝孢子组孕鼠胚胎的神经管畸形发生率显著低于实验对照组。神经管神经上皮细胞表达nestin的百分率也明显大于实验对照组。实验对照组孕鼠胚胎的神经管细胞在G0/G1期的比例则显著高于灵芝孢子组,但是实验对照组神经管细胞S期的比例低,低于灵芝孢子组形态正常的胚胎神经管细胞。RT-PCR检测发现,灵芝孢子组孕鼠胚胎神经管细胞能够正常转录Cdk4 mRNA,而实验对照组则低转录Cdk4 mRNA。结论灵芝孢子能够减少维甲酸诱导的妊娠小鼠孕期E10.5d胚胎神经管畸形的发生。
Objective To investigate whether Ganoderma lucidum spores can promote the induction of retinoid acid, but the neuronal epithelial cells in embryonic neural tube arrested in G0/G1 phase re-enter the cell cycle, continue to proliferate and differentiate, and reduce the occurrence of neural tube defects. METHODS: The experimental control group and the ganoderma lucidum spore group mice received a one-time gastric feeding of all-trans retinoic acid at the time of 7.75 days of pregnancy embryo (E), resulting in neural tube defects in the embryos of the two groups of pregnant mice. Then, the Ganoderma lucidum spore group was given a stomach-fed Ganoderma spore solution. Embryos of two groups of pregnant rats were taken at E10.5d during pregnancy, and the incidence of neural tube defects in embryos was counted. Immunofluorescence staining and flow cytometry were used to detect the expression of nestin in embryonic neural tube neuroepithelium. Quantitative fluorescent staining and RT-PCR were used to detect the transcription of embryonic neural tube dependent cyclin kinase 2 (Cdk2) mRNA and Cdk4 mRNA. Results The incidence of neural tube defects in the embryo of the Ganoderma lucidum spore group was significantly lower than that of the experimental control group. The percentage of nestin expressed in neural tube epithelial cells was also significantly greater than that of the experimental control group. The proportion of neural tube cells in the experimental control embryos at the G0/G1 phase was significantly higher than that of the Ganoderma spores group, but the proportion of S phase in the neural tube cells was lower in the experimental control group than that in the normal control embryonic neural tube cells of the Ganoderma lucidum spore group. . RT-PCR showed that the neural tube cells of embryonic rat embryos in the ganoderma lucidum spore group were able to normally transcribe Cdk4 mRNA, whereas in the experimental control group, Cdk4 mRNA was lowly transcribed. Conclusion Ganoderma lucidum spores can reduce the incidence of embryonic neural tube defects in embryonic mice induced by retinoic acid during pregnancy E10.5d.