体外培养大鼠脑皮质神经元离心损伤模型的建立

Establishment of contical neuron injury model with in vitro culture in rat

目的建立体外培养大鼠脑皮质神经元离心损伤模型,观察离心损伤后神经元的形态学变化和培养液中乳酸脱氢酶(LDH)的含量变化。方法把培养神经元的六孔培养板的孔内,置入与孔大小相当的200目钢网,并在钢网上放置6g重物,通过变速离心培养板,造成神经元损伤的作用。使用倒置相差显微镜,电镜观察损伤后神经元的形态改变,并测量培养液中乳酸脱氢酶 (LDH)的含量变化。结果光镜可见神经元破碎、轴索肿胀、轴索球、轴索部分崩解。电镜可见细胞骨架发生变化。从神经元损伤15min后,培养上清液中LDH活性增高。损伤组LDH升高有两个时相,第一峰在30min出现,第二峰在72h出现,损伤后30min,24h,48h,72h,损伤组与对照组同时间相比,LDH含量均有显著统计学意义(P<0．01),第一峰值可能是原发性损伤造成的细胞膜破损,LDH释放量升高引起的,第二峰值可能是继发性损伤导致细胞膜破损,LDH漏出所引起的。结论实验建立的神经元损伤模型操作简便,能较好地模拟脑外伤。

Objective A traumatic neuron injury model emphasizing on the axonal injury was created. Method By menas of centrifugating culture plate with vital neuron in vitro, the shape of axons was ohserved with microscope, and the microtube and microfilament of the axons were observed with electron microscope. Meanwhile lactate dehydrogenase （LDH） was measured in culture medium at different stages, and compared with control group, then analysed stastically. Result Axonal swelling, axonal bulb, disruption of axon and neuron were observed with microscope, a decrease in the number of microtube and microfilament of the axons were observed with electron microscope. In injury group, LDH showed difference with control （P 〈 0.01） . Conclusion The results suggested that the traumatic neuron injury model was created rationally and practically.