预先服用灵芝孢子降低维甲酸诱导的孕鼠胚胎神经管畸形发生率  

Pre-administration of Ganoderma lucidum spore reduces incidence of neural tube defects induced by retinoic acid in pregnant mice

作　　者：张伟 曾园山[1] 熊轶[1] 陈穗君[1] 钟志强[1] 

机构地区：[1]中山大学中山医学院组织胚胎学教研室神经科学研究室,广东广州510080

出　　处：《中西医结合学报》2006年第4期368-373,共6页Journal of Chinese Integrative Medicine

摘　　要：目的探讨预先服用灵芝孢子能否降低维甲酸诱导的胚胎神经管畸形的发生率。方法将胚胎(embryo,E)0d孕鼠随机分为正常对照组、溶剂组、模型组和灵芝孢子组,每组5只。灵芝孢子组孕鼠于E0d起即给予胃饲灵芝孢子溶液,溶剂组孕鼠胃饲等量溶剂。至E7.75d时,模型组和灵芝孢子组孕鼠一次性胃饲全反式维甲酸。E10.5d时取孕鼠胚胎,计算神经管畸形发生率及测量胚胎顶臀距。采用免疫荧光组织化学染色和流式细胞术检测胚胎神经管神经上皮细胞巢蛋白的表达及细胞周期分布,采用DNA定量荧光染色及RT-PCR技术检测胚胎神经管细胞周期素依赖性蛋白激酶2(cyclin-dependentkinase2,Cdk2)和Cdk4mRNA的转录水平。结果孕鼠胚胎神经管畸形发生率模型组为79.41%,灵芝孢子组为21.67%;顶臀距模型组为(3.62±1.27)mm,灵芝孢子组为(5.84±0.92)mm。胚胎神经管神经上皮细胞巢蛋白表达阳性率模型组为32.44%,灵芝孢子组为77.65%。模型组孕鼠胚胎神经管神经上皮细胞G0/G1期比例为82.80%,灵芝孢子组为53.74%。灵芝孢子组Cdk4mRNA转录水平高于正常对照组和溶剂组,但未检测到Cdk2mRNA。结论预先服用灵芝孢子能够降低维甲酸诱导的孕鼠胚胎神经管畸形的发生率。Objective： To explore whether pre-administration of Ganoderma lucidum spore （GASP） can reduce incidence of neural tube defects （NTDs） induced by all-trans retinoic acid （ATRA） in pregnant mice. Methods： Twenty pregnant mice were randomly divided into four groups： normal control group, solvent- treated group, ATRA-induced group, and GASP-treated plus ATRA-induced group. GASP solution, which was prepared with solvent （sodium carboxymethyl cellulose）, was fed to the pregnant mice in the GASP-treated plus ATRA-induced group twice a day from embryo （E） 0 d to E10.5 d. The same dose of solvent was given to the pregnant mice in the solvent-treated group. At E7.75 d, ATRA （50 mg/kg） was given to the pregnant mice in both ATRA-induced group and GASP-treated plus ATRA-induced group for single time. Embryos were sampled from pregnant mice at E10.5 d. Then the incidence rate of NTDs in mouse embryo was calculated and the crown-rump length of mouse embryo was measured. The positive rate of nestin expression and the distribution of cell cycle of embryonic neural tube neuroepithelial cells were detected by histochemical staining technique and flow cytometry respectively. Reverse transcription-polymerase chain reaction method was used to detect the gene expressions of cyclin-dependent protein kinase 2 （Cdk2） and Cdk4 mRNAs. Results： The incidence rates of NTDs in mouse embryos in the ATRA-induced group and the GASP-treated plus ATRA-induced group were 79.41% and 21.67% respectively, while the crown-rump length of mouse embryos in these two groups were （3.62 ±1.27） mm and （5.84±0.92） mm respectively. The positive rate of nestin expression in embryonic neural tube neuroepithelial cells of mouse embryo at El0.5 d in the ATRA-induced group was 32.44%, while that in the GASP-treated plus ATRA-induced group was 77.65%. The cell cycle of embryonic neural tube neuroepithelial cells was obviously arrested at Go/G, phase in the ATRA-induced group as compared with that in the GASP-treated plus

关 键 词：神经管畸形 灵芝孢子 维甲酸 巢蛋白 细胞周期素依赖性蛋白激酶 

分 类 号：R285.5[医药卫生—中药学]