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目的 观察胶质细胞源性神经营养因子 (GDNF)及单纯疱疹病毒载体介导的 GDNF(HSV- GDNF)对体外培养的胎鼠脊髓运动神经元在划痕损伤后凋亡的影响。 方法 对培养 12 d神经元行划痕损伤 ,并将其分成 4组 (无血清对照组、血清组、HSV- GDNF组和 GDNF组 ) ,给予不同培养液 ,定期观察各组的运动神经元存活数。分别于划痕损伤第 4d和第 7d时对神经元作 TU NEL 染色 ,检测运动神经元凋亡数 ,并在图像分析仪上对凋亡神经元作平均光密度的色谱分析。 结果 各组内运动神经元存活数与培养时间成反比。从对照组、HSV- GDNF组到 GDNF组 ,运动神经元凋亡数和凋亡神经元的平均光密度均依次减少 ,但对照组和血清组、GDNF组和 HSV-GDNF组间的运动神经元凋亡数以及凋亡神经元平均光密度均无显著差异。 结论 GDNF和 HSV- GDNF能挽救生长发育过程中受损的脊髓运动神经元的凋亡 ,对体外培养的受损脊髓运动神经元具有一定的保护作用。
Objective To investigate the effects of GDNF and HSV-GDNF on the apoptosis of fetal rat spinal cord motor neurons cultured in vitro. Methods Scratch injury of neurons cultured for 12 days was divided into 4 groups (serum-free control group, serum group, HSV-GDNF group and GDNF group). Different culture mediums were given and the survival of motor neurons in each group was observed regularly number. The neurons were stained with TUNEL on the 4th and 7th day respectively. The number of apoptosis of motor neurons was detected. The average optical density of apoptotic neurons was analyzed by the image analyzer. Results The survival of motor neurons in each group was inversely proportional to the culture time. From the control group, the HSV-GDNF group to the GDNF group, the number of apoptotic neurons and the average optical density of apoptotic neurons decreased in turn, but the motor neurons between the control group and the serum group, the GDNF group and the HSV-GDNF group There was no significant difference in apoptotic number and apoptotic neuron average optical density. Conclusion GDNF and HSV-GDNF can rescue the apoptosis of injured spinal motor neurons during growth and development, and have protective effects on injured spinal cord motor neurons cultured in vitro.