石杉碱甲对体外培养大鼠脊髓片前角运动神经元损伤的保护作用  

Protection of huperzine A against motor neuron injury in cultured rat spinal cord slice

作　　者：刘卫刚[1] 肖向建[1] 李敏[1] 马征[1] 李春岩[1] 

机构地区：[1]河北医科大学第二医院神经内科,河北石家庄050000

出　　处：《中国新药与临床杂志》2005年第12期933-937,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：河北省自然科学基金资助项目(303487)

摘　　要：目的:利用选择性运动神经元死亡的脊髓薄片培养模型,观察石杉碱甲对运动神经元的保护作用。方法:乳大鼠腰段脊髓组织薄片分为正常对照组、模型组和石杉碱甲1,10,100μmol·L-15组。用倒置显微镜观察脊髓片形态变化,SMI32免疫组化染色对腹角α运动神经元记数,透射电镜观察α运动神经元的超微结构并测定培养液中乳酸脱氢酶(LDH)的含量。结果:石杉碱甲100μmol·L-1组脊髓片在体外生长良好,形态与正常对照组相近;石杉碱甲1,10μmol·L-1组脊髓片生长不良,形态与模型组相近。与正常对照组相比,模型组脊髓片前角SMI32阳性α运动神经元数减少(P<0.05),培养液中LDH含量增加(P<0.05)。与模型组相比,石杉碱甲100μmol·L-1组脊髓片前角α运动神经元数增加(P<0.05),胞浆内空泡明显减少,培养液中LDH含量减低(P<0.05)。结论:高浓度石杉碱甲能改善乳大鼠脊髓片前角α运动神经元生长,减少其空泡形成及死亡,并降低LDH的释放。AIM： To observe the protection of huperzine A on motor neuron injury in cultured spinal cord slice model of selective motor neuron death. METHODS： The lumbar spinal cord slices from rat pups were divided into five groups randomly, normal control group, model group and huperzine A 1, 10, 100 μmol·L^-1 group. The effects of drug were observed with inverted microscope for inspecting the morphologic change of spinal cord slice, SMI-32 immunohistochemical staining for calculating ventral angle ct motor neuron, assay of lactate dehydrogenase （LDH） in culture and supermicro structure of ct motor neurons through transmission electron microscope. RESULTS： The spinal cord slices of huperzine A 100 μmol·L^-1 group had better growth and showed the similar change with the slices of normal control group; the slices of huperzine A 1, 10 μmol·L^-1 groups had poor growth and with similar morphology to the slices of model group. After 4 wk in vitro, the number of SMI-32 positive α motor neurons in spinal cord slice decreased （ P 〈 0.05 ） and increase of LDH in culture medium （ P 〈 0.05 ） of the model group vs normal control group. Comparing to model group, increase number of α motor neurons （ P 〈 0.05 ）, decrease of LDH in culture medium （ P 〈 0.05 ） and disapperance of cytoplasmic vacuoles were shown apparently in huperzine A 100 μmol·L^-1 group. CONCLUSION： High concentration of huperzine A can improve the condition of motor neuron growth, reduce the formation and death of cytoplasmic vacuoles and decrease the release of LDH.

关 键 词：石杉碱甲 肌萎缩侧索硬化 脊髓 谷氨酸 细胞培养 苏-羟天冬氨酸 

分 类 号：R744.8[医药卫生—神经病学与精神病学]