大鼠背根神经节加压培养细胞模型的建立

目的建立大鼠背根神经节（DRG）细胞受压模型，观察在恒定压力下培养24h和48h后神经元形态和活性的变化，评价这种神经元压力培养模型的优缺点。方法将培养的新生大鼠DRG细胞随机分为正常对照组、受压24h组、受压48h组，在80mmHg压力的密闭环境中培养相应时间后观察各组细胞在荧光显微镜和电镜下形态学特点，并用MTT法分析各组神经元细胞生长活性的改变。结果加压培养后，荧光显微镜下DRG神经元形态未发生明显变化，超微结构示48h组和24h组较正常组线粒体肿胀明显，粗面内质网缩短、变少，游离核糖体增多，部分核固缩，神经元代谢水平下降，MTT法比较加压前、后各组细胞活性的差异无统计学意义。结论加压培养未使细胞活性发生显著改变，此模型可观察机械压力单一因素对神经元的影响，有望成为一种在亚细胞和分子水平研究机械压力对DRG神经元的影响的有效细胞模型。

A novel compressing cultivation model of rat dorsal root ganglion neurons in vitro

Objective To establish a compressing model of rat dorsal root ganglion neurons cultivation in vitro and observe the changes in appearance and activity in cultured rat dorsal root ganglion ( DRG) neurons under constant pressure(80 mmHg)and different time intervals,so as to evaluate the advantages and disadvantages of the neuron culture model. Methods According to the pressure exerting time,the cultured DRG cells was assigned to three groups :24 h group,48 h group and normal control group,the changes of appearance and activity were demonstrated by fluorescent microscope, transmission electron microscopy and MTT assay. Results There was no significant difference among groups under fluorescent microscope. In the 24 h group and 48 h group,compared with control group,the electron microscopy results of ultra-structure suggested that the high-pressure cultivation environment would influence the metabolism of neurons: the number of the rough endoplasmic reticulum decreased, and its length was shorten, mitochondrion swelled and some nucleolus concentrated, electron microscope also showed the increment of free ribosome. No significant activity changes was detected by MTT method among groups (P >0.05). Conclusion The novel compressing model of DRG neuron could avoid the defect of the models in vivo, and provide the convenience of observing the function of single pressure factor exactly. This model would be an effective tool for the studies of the effect of mechanical compression on DRG neurons.