GDNF及HSV-GDNF对体外培养大鼠脊髓运动神经元受损后凋亡的影响

EFFECTS OF BOTH GDNF AND HSV-GDNF ON APOPTOSIS OF INJURED SPINAL CORD MOTOR NEURONS in vitro

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 study effects of GDNF and HSV GDNF on apoptosis of spinal cord motoneurons after scratch injury in vitro. Methods In the period of culture cell,motor neurons were periodically observed and counted.Scratch injury was executed on culturing 12th day,in the same time,cultured neurons were divided into 4 groups,and each group was given corresponding medium(medium serum free control group,serum group,HSV GDNF group,GDNF group).On the 4th and 7th day after scratch injury,TUNEL staining was respectively performed,and the number and the mean densities of apoptotic motoneurons were observed. Results The number of living motoneurons was in inverse proportion to time of scratch injury in each group.The number of apoptotic motoneurons from control group,HSV GDNF group to GDNF group was successively decreased as well as the mean densities of apoptotic motoneurons on the 4th and 7th day after scratch injury.Furthermore,the effects of groups with serum were no better than those of medium serum free groups,in the same time,difference was not obviously in HSV GDNF group and GDNF group. Conclusion GDNF and HSV GDNF can decrease apoptosis of injured motoneurons in vitro .It suggests that GDNF and HSV GDNF might play an important role in the growth and development of motor neurons.