人参皂甙Rd对脊髓运动神经元抗谷氨酸损伤的作用观察  

Neuroprotective effect of Ginsenoside Rd on spinal cord motoneurons from glutamate-induced injury in vitro

作　　者：马丽[1] 桑韩飞[2] 刘永勤[1] 

机构地区：[1]海军总医院麻醉科,北京100048 [2]第四军医大学西京医院麻醉科,西安710032

出　　处：《解放军医学杂志》2012年第10期955-957,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30600840)~~

摘　　要：目的探讨人参皂甙Rd对体外培养的脊髓运动神经元抗谷氨酸损伤的作用。方法分离胎龄15d的SD大鼠的脊髓运动神经元,培养6d后分为4组:对照组,培养基中不加药液;谷氨酸(Glu)组,培养基中加入300μmol/L谷氨酸;人参皂甙Rd+谷氨酸(GSRd+Glu)组,培养基中预先加入10μmol/L人参皂甙Rd孵育30min,再加入300μmol/L谷氨酸;丙二醇+谷氨酸(Vehicle+Glu)组,培养基中预先加入等容量丙二醇孵育30min,再加入300μmol/L谷氨酸。继续培养24h,相差显微镜下观察神经元形态及生长状况,进行细胞计数,测定神经元存活率及培养上清液中乳酸脱氢酶(LDH)活性。结果 Glu组存活的脊髓运动神经元数目少,偶有突起形成,细胞受损严重;GSRd+Glu组脊髓运动神经元数目明显增多(P<0.05),突起较多,只少数死亡细胞;Vehicle+Glu组神经元形态与Glu组相似。Glu组及Vehicle+Glu组脊髓运动神经元存活率分别为22.7%±2.3%、23.6%±2.1%,明显低于对照组(100%±0%,P<0.05)及GSRd+Glu组(58.6%±2.3%,P<0.05),而Glu组与Vehicle+Glu组比较、对照组与GSRd+Glu组比较差异均无统计学意义(P>0.05)。Glu组及Vehicle+Glu组培养上清中LDH活性分别为142.50±3.55、139.20±2.75U/L,明显高于对照组(42.3±3.6U/L)及GSRd+Glu组(84.2±2.5U/L,P<0.05),而Glu组与Vehicle+Glu组比较、对照组与GSRd+Glu组比较差异均无统计学意义(P>0.05)。结论人参皂甙Rd对脊髓运动神经元抗谷氨酸损伤具有保护作用。Objective To investigate the neuroprotective effect of ginsenoside Rd （GSRd） on glutamate-induced spinal cord motoneuron injury in vitro. Methods The spinal cord motoneurons obtained from 1S-day SD rat embryos were cultured for 6 days, and then randomly assigned into four groups as follows. Control： no treatment; Glu： glutamate 300p^mol/L was added to the spinal cord neurons; GSRd±Glu： the spinal cord neurons was pretreated with 10p, mol/L GSRd for 30min, and then glutamate 300~mol/L was added to the spinal cord neurons; Vehicle±GIu： the spinal cord neurons was pretreated with propylene glycol at the same volume as that used for GSRd, and then glutamate 300~mol/L was added to the spinal cord neurons. After the cultures were incubated with glutamate for 24h, the motor neurons were counted and observed for Changes in morphology and state of growth under phase-contrast microscope; the neuron viability was determined by methyl thiazolyl terazolium assay （MTf）; the activity of lactate dehydrogenase （LDH） leaked into the culture medium was also measured. Results There were less living spinal motor neurons and neuronal processes in Glu group than in GSRd±Glu group. The nervous cells were severely damaged in Glu group, while the number of dead cells was very small in GSRd±Glu group. The neuronal morphology in Vehicle±Glu group was similar to that in Glu group. The survival rates of spinal motor neurons were 22.7% ± 2.3% and 23.6% ± 2.1% in Glu group and Vehicle±Glu group, respectively, and they were significantly lower than those in control group （100% ± 0%, P〈0.05） and GSRd±Glu group （58.6% ± 2.3%, P〈0.0S）, while there was no significant difference between Glu group and Vehicle±Glu group and between control and GSRd±Glu groups （P〉0.05）. The LDH activity in the culture supernatant was 142.50 ± 3.55 and 139.20 ± 2.75U/ L respectively in Glu and Vehicle±Glu, which were significantly higher than that in control group （42.3 ± 3.6U/L, P〈0.0S） and GSRd±Glu （84.2 �

关 键 词：脊髓 再灌注损伤 人参皂甙 运动神经元 谷氨酸 

分 类 号：R651.21[医药卫生—外科学]