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从大鼠胚胎肢芽中提取生物活性物质,研究其对体外培养的胚鼠背根神经节感觉神经元的作用。方法:剖腹取孕10～15天的SD大鼠胚胎,将胚胎肢芽的提取物加至胚鼠背根神经节感觉神经元的体外培养基中,同单纯培养基培养作对照。接种后,采用相差倒置显微镜、电镜、台盼蓝染色计数和乙酰胆碱脂酶(AchE)定量测定等技术观察培养细胞的生长、功能、超微结构和成活率的变化。结果:实验组细胞的成活率和轴突的长度明显优于对照组(P<0.01);AchE的含量明显多于对照组(P<0.01)。电镜下实验组线粒体、核糖体和高尔基复合体等细胞器的超微结构和功能状态也明显优于对照组。结论:胚胎肢芽提取物(ELBE)具有较强的维持神经元存活,促进轴突生长的神经营养因子(NTFs)活性。
Bioactive substances were extracted from rat embryonic limb buds to study their effect on sensory neurons of dorsal root ganglion in vitro. METHODS: Sprague-Dawley (SD) rats embryos 10 to 15 days after caesarean section were implanted. The extracts of embryonic limb buds were added to the in vitro culture medium of sensory neurons of dorsal root ganglion of embryonic rats, and cultured with simple culture medium. After inoculation, the changes of growth, function, ultrastructure and survival rate of cultured cells were observed by inverted phase contrast microscope, electron microscope, trypan blue staining and quantitative determination of acetylcholinesterase (AchE). Results: The survival rate and axon length of the experimental group were significantly better than those of the control group (P <0.01). The content of AchE in the experimental group was more than that of the control group (P <0.01). The ultrastructure and functional status of organelles such as mitochondria, ribosome and Golgi complex in the experimental group under electron microscope were also significantly better than those in the control group. Conclusion: Embryonic limb bud extract (ELBE) has strong neurotrophic factor (NTFs) activity to maintain neuronal survival and promote axonal growth.